Compositions and methods for the therapy and diagnosis of breast cancer

ABSTRACT

Compositions and methods for the therapy and diagnosis of cancer, particularly breast cancer, are disclosed. Illustrative compositions comprise one or more breast tumor polypeptides, immunogenic portions thereof, polynucleotides that encode such polypeptides, antigen presenting cell that expresses such polypeptides, and T cells that are specific for cells expressing such polypeptides. The disclosed compositions are useful, for example, in the diagnosis, prevention and/or treatment of diseases, particularly breast cancer.

BACKGROUND OF THE INVENTION

[0001] 1. Field of the Invention

[0002] The present invention relates generally to therapy and diagnosisof cancer, such as breast cancer. The invention is more specificallyrelated to polypeptides, comprising at least a portion of a breast tumorprotein, and to polynucleotides encoding such polypeptides. Suchpolypeptides and polynucleotides are useful in pharmaceuticalcompositions, e.g., vaccines, and other compositions for the diagnosisand treatment of breast cancer.

[0003] 2. Description of the Related Art

[0004] Cancer is a significant health problem throughout the world.Although advances have been made in detection and therapy of cancer, novaccine or other universally successful method for prevention and/ortreatment is currently available. Current therapies, which are generallybased on a combination of chemotherapy or surgery and radiation,continue to prove inadequate in many patients.

[0005] Breast cancer is a significant health problem for women in theUnited States and throughout the world. Although advances have been madein detection and treatment of the disease, breast cancer remains thesecond leading cause of cancer-related deaths in women, affecting morethan 180,000 women in the United States each year. For women in NorthAmerica, the life-time odds of getting breast cancer are now one ineight.

[0006] No vaccine or other universally successful method for theprevention or treatment of breast cancer is currently available.Management of the disease currently relies on a combination of earlydiagnosis (through routine breast screening procedures) and aggressivetreatment, which may include one or more of a variety of treatments suchas surgery, radiotherapy, chemotherapy and hormone therapy. The courseof treatment for a particular breast cancer is often selected based on avariety of prognostic parameters, including an analysis of specifictumor markers. See, e.g., Porter-Jordan and Lippman, Breast Cancer8:73-100 (1994). However, the use of established markers often leads toa result that is difficult to interpret, and the high mortality observedin breast cancer patients indicates that improvements are needed in thetreatment, diagnosis and prevention of the disease.

[0007] In spite of considerable research into therapies for these andother cancers, breast cancer remains difficult to diagnose and treateffectively. Accordingly, there is a need in the art for improvedmethods for detecting and treating such cancers. The present inventionfulfills these needs and further provides other related advantages.

SUMMARY OF THE INVENTION

[0008] In one aspect, the present invention provides polynucleotidecompositions comprising a sequence selected from the group consistingof:

[0009] (a) sequences provided in SEQ ID NO: 1, 3-86, 142-298, 301-303,307, 313, 314, 316, 317, 323, 325, 327-330, 335, 339, and 341-344;

[0010] (b) complements of the sequences provided in SEQ ID NO: 1, 3-86,142-298, 301-303, 307, 313, 314, 316, 317, 323, 325, 327-330, 335, 339,and 341-344;

[0011] (c) sequences consisting of at least 20, 25, 30, 35, 40, 45, 50,75 and 100 contiguous residues of a sequence provided in SEQ ID NO: 1,3-86, 142-298, 301-303, 307, 313, 314, 316, 317, 323, 325, 327-330, 335,339, and 341-344;

[0012] (d) sequences that hybridize to a sequence provided in SEQ ID NO:1, 3-86, 142-298, 301-303, 307, 313, 314, 316, 317, 323, 325, 327-330,335, 339, and 341-344, under moderate or highly stringent conditions;

[0013] (e) sequences having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%,98% or 99% identity to a sequence of SEQ ID NO: 1, 3-86, 142-298,301-303, 307, 313, 314, 316, 317, 323, 325, 327-330, 335, 339, and341-344;

[0014] (f) degenerate variants of a sequence provided in SEQ ID NO: 1,3-86, 142-298, 301-303, 307, 313, 314, 316, 317, 323, 325, 327-330, 335,339, and 341-344.

[0015] In one preferred embodiment, the polynucleotide compositions ofthe invention are expressed in at least about 20%, more preferably in atleast about 30%, and most preferably in at least about 50% of breasttumors samples tested, at a level that is at least about 2-fold,preferably at least about 5-fold, and most preferably at least about10-fold higher than that for normal tissues.

[0016] The present invention, in another aspect, provides polypeptidecompositions comprising an amino acid sequence that is encoded by apolynucleotide sequence described above.

[0017] The present invention further provides polypeptide compositionscomprising an amino acid sequence selected from the group consisting ofsequences recited in SEQ ID NO: 131-140, 299, 300, 304-306, 308-312,315, 318, 324, 326, 331-334, 336, 340, and 345-428.

[0018] In certain preferred embodiments, the polypeptides and/orpolynucleotides of the present invention are immunogenic, i.e., they arecapable of eliciting an immune response, particularly a humoral and/orcellular immune response, as further described herein.

[0019] The present invention further provides fragments, variants and/orderivatives of the disclosed polypeptide and/or polynucleotidesequences, wherein the fragments, variants and/or derivatives preferablyhave a level of immunogenic activity of at least about 50%, preferablyat least about 70% and more preferably at least about 90% of the levelof immunogenic activity of a polypeptide sequence set forth in SEQ IDNO: 131-140, 299, 300, 304-306, 308-312, 315, 318, 324, 326, 331-334,336, 340, and 345-428 or a polypeptide sequence encoded by apolynucleotide sequence set forth in SEQ ID NO: 1, 3-86, 142-298,301-303, 307, 313, 314, 316, 317, 323, 325, 327-330, 335, 339, and341-344.

[0020] The present invention further provides polynucleotides thatencode a polypeptide described above, expression vectors comprising suchpolynucleotides and host cells transformed or transfected with suchexpression vectors.

[0021] Within other aspects, the present invention providespharmaceutical compositions comprising a polypeptide or polynucleotideas described above and a physiologically acceptable carrier.

[0022] Within a related aspect of the present invention, thepharmaceutical compositions, e.g., vaccine compositions, are providedfor prophylactic or therapeutic applications. Such compositionsgenerally comprise an immunogenic polypeptide or polynucleotide of theinvention and an immunostimulant, such as an adjuvant.

[0023] The present invention further provides pharmaceuticalcompositions that comprise: (a) an antibody or antigen-binding fragmentthereof that specifically binds to a polypeptide of the presentinvention, or a fragment thereof; and (b) a physiologically acceptablecarrier.

[0024] Within further aspects, the present invention providespharmaceutical compositions comprising: (a) an antigen presenting cellthat expresses a polypeptide as described above and (b) apharmaceutically acceptable carrier or excipient. Illustrative antigenpresenting cells include dendritic cells, macrophages, monocytes,fibroblasts and B cells.

[0025] Within related aspects, pharmaceutical compositions are providedthat comprise: (a) an antigen presenting cell that expresses apolypeptide as described above and (b) an immunostimulant.

[0026] The present invention further provides, in other aspects, fusionproteins that comprise at least one polypeptide as described above, aswell as polynucleotides encoding such fusion proteins, typically in theform of pharmaceutical compositions, e.g., vaccine compositions,comprising a physiologically acceptable carrier and/or animmunostimulant. The fusions proteins may comprise multiple immunogenicpolypeptides or portions/variants thereof, as described herein, and mayfurther comprise one or more polypeptide segments for facilitating theexpression, purification and/or immunogenicity of the polypeptide(s).

[0027] Within further aspects, the present invention provides methodsfor stimulating an immune response in a patient, preferably a T cellresponse in a human patient, comprising administering a pharmaceuticalcomposition described herein. The patient may be afflicted with breastcancer, in which case the methods provide treatment for the disease, orpatient considered at risk for such a disease may be treatedprophylactically.

[0028] Within further aspects, the present invention provides methodsfor inhibiting the development of a cancer in a patient, comprisingadministering to a patient a pharmaceutical composition as recitedabove. The patient may be afflicted with breast cancer, in which casethe methods provide treatment for the disease, or patient considered atrisk for such a disease may be treated prophylactically.

[0029] The present invention further provides, within other aspects,methods for removing tumor cells from a biological sample, comprisingcontacting a biological sample with T cells that specifically react witha polypeptide of the present invention, wherein the step of contactingis performed under conditions and for a time sufficient to permit theremoval of cells expressing the protein from the sample.

[0030] Within related aspects, methods are provided for inhibiting thedevelopment of a cancer in a patient, comprising administering to apatient a biological sample treated as described above.

[0031] Methods are further provided, within other aspects, forstimulating and/or expanding T cells specific for a polypeptide of thepresent invention, comprising contacting T cells with one or more of:(i) a polypeptide as described above; (ii) a polynucleotide encodingsuch a polypeptide; and/or (iii) an antigen presenting cell thatexpresses such a polypeptide; under conditions and for a time sufficientto permit the stimulation and/or expansion of T cells. Isolated T cellpopulations comprising T cells prepared as described above are alsoprovided.

[0032] Within further aspects, the present invention provides methodsfor inhibiting the development of a cancer in a patient, comprisingadministering to a patient an effective amount of a T cell population asdescribed above.

[0033] The present invention further provides methods for inhibiting thedevelopment of a cancer in a patient, comprising the steps of: (a)incubating CD4⁺ and/or CD8⁺ T cells isolated from a patient with one ormore of: (i) a polypeptide comprising at least an immunogenic portion ofpolypeptide disclosed herein; (ii) a polynucleotide encoding such apolypeptide; and (iii) an antigen-presenting cell that expressed such apolypeptide; and (b) administering to the patient an effective amount ofthe proliferated T cells, and thereby inhibiting the development of acancer in the patient. Proliferated cells may, but need not, be clonedprior to administration to the patient.

[0034] Within further aspects, the present invention provides methodsfor determining the presence or absence of a cancer, preferably a breastcancer, in a patient comprising: (a) contacting a biological sampleobtained from a patient with a binding agent that binds to a polypeptideas recited above; (b) detecting in the sample an amount of polypeptidethat binds to the binding agent; and (c) comparing the amount ofpolypeptide with a predetermined cut-off value, and therefromdetermining the presence or absence of a cancer in the patient. Withinpreferred embodiments, the binding agent is an antibody, more preferablya monoclonal antibody.

[0035] The present invention also provides, within other aspects,methods for monitoring the progression of a cancer in a patient. Suchmethods comprise the steps of: (a) contacting a biological sampleobtained from a patient at a first point in time with a binding agentthat binds to a polypeptide as recited above; (b) detecting in thesample an amount of polypeptide that binds to the binding agent; (c)repeating steps (a) and (b) using a biological sample obtained from thepatient at a subsequent point in time; and (d) comparing the amount ofpolypeptide detected in step (c) with the amount detected in step (b)and therefrom monitoring the progression of the cancer in the patient.

[0036] The present invention further provides, within other aspects,methods for determining the presence or absence of a cancer in apatient, comprising the steps of: (a) contacting a biological sample,e.g., tumor sample, serum sample, etc., obtained from a patient with anoligonucleotide that hybridizes to a polynucleotide that encodes apolypeptide of the present invention; (b) detecting in the sample alevel of a polynucleotide, preferably mRNA, that hybridizes to theoligonucleotide; and (c) comparing the level of polynucleotide thathybridizes to the oligonucleotide with a predetermined cut-off value,and therefrom determining the presence or absence of a cancer in thepatient. Within certain embodiments, the amount of mRNA is detected viapolymerase chain reaction using, for example, at least oneoligonucleotide primer that hybridizes to a polynucleotide encoding apolypeptide as recited above, or a complement of such a polynucleotide.Within other embodiments, the amount of mRNA is detected using ahybridization technique, employing an oligonucleotide probe thathybridizes to a polynucleotide that encodes a polypeptide as recitedabove, or a complement of such a polynucleotide.

[0037] In related aspects, methods are provided for monitoring theprogression of a cancer in a patient, comprising the steps of: (a)contacting a biological sample obtained from a patient with anoligonucleotide that hybridizes to a polynucleotide that encodes apolypeptide of the present invention; (b) detecting in the sample anamount of a polynucleotide that hybridizes to the oligonucleotide; (c)repeating steps (a) and (b) using a biological sample obtained from thepatient at a subsequent point in time; and (d) comparing the amount ofpolynucleotide detected in step (c) with the amount detected in step (b)and therefrom monitoring the progression of the cancer in the patient.

[0038] Within further aspects, the present invention providesantibodies, such as monoclonal antibodies, that bind to a polypeptide asdescribed above, as well as diagnostic kits comprising such antibodies.Diagnostic kits comprising one or more oligonucleotide probes or primersas described above are also provided.

[0039] These and other aspects of the present invention will becomeapparent upon reference to the following detailed description andattached drawings. All references disclosed herein are herebyincorporated by reference in their entirety as if each was incorporatedindividually.

BRIEF DESCRIPTION OF THE DRAWINGS

[0040]FIG. 1 shows the differential display PCR products, separated bygel electrophoresis, obtained from cDNA prepared from normal breasttissue (lanes 1 and 2) and from cDNA prepared from breast tumor tissuefrom the same patient (lanes 3 and 4). The arrow indicates the bandcorresponding to B18Ag1.

[0041]FIG. 2 is a northern blot comparing the level of B18Ag1 mRNA inbreast tumor tissue (lane 1) with the level in normal breast tissue.

[0042]FIG. 3 shows the level of B18Ag1 mRNA in breast tumor tissuecompared to that in various normal and non-breast tumor tissues asdetermined by RNase protection assays.

[0043]FIG. 4 is a genomic clone map showing the location of additionalretroviral sequences obtained from ends of XbaI restriction digests(provided in SEQ ID NO: 3-SEQ ID NO: 10) relative to B18Ag1.

[0044]FIGS. 5A and 5B show the sequencing strategy, genomic organizationand predicted open reading frame for the retroviral element containingB18Ag1.

[0045]FIG. 6 shows the nucleotide sequence of the representative breasttumor-specific cDNA B18Ag1.

[0046]FIG. 7 shows the nucleotide sequence of the representative breasttumor-specific cDNA B17Ag1.

[0047]FIG. 8 shows the nucleotide sequence of the representative breasttumor-specific cDNA B17Ag2.

[0048]FIG. 9 shows the nucleotide sequence of the representative breasttumor-specific cDNA B13Ag2a.

[0049]FIG. 10 shows the nucleotide sequence of the representative breasttumor-specific cDNA B13Ag1b.

[0050]FIG. 11 shows the nucleotide sequence of the representative breasttumor-specific cDNA B13Ag1a.

[0051]FIG. 12 shows the nucleotide sequence of the representative breasttumor-specific cDNA B11Ag1.

[0052]FIG. 13 shows the nucleotide sequence of the representative breasttumor-specific cDNA B3CA3c.

[0053]FIG. 14 shows the nucleotide sequence of the representative breasttumor-specific cDNA B9CG1.

[0054]FIG. 15 shows the nucleotide sequence of the representative breasttumor-specific cDNA B9CG3.

[0055]FIG. 16 shows the nucleotide sequence of the representative breasttumor-specific cDNA B2CA2.

[0056]FIG. 17 shows the nucleotide sequence of the representative breasttumor-specific cDNA B3CA1.

[0057]FIG. 18 shows the nucleotide sequence of the representative breasttumor-specific cDNA B3CA2.

[0058]FIG. 19 shows the nucleotide sequence of the representative breasttumor-specific cDNA B3CA3.

[0059]FIG. 20 shows the nucleotide sequence of the representative breasttumor-specific cDNA B4CA1.

[0060]FIG. 21A depicts RT-PCR analysis of breast tumor genes in breasttumor tissues (lanes 1-8) and normal breast tissues (lanes 9-13) and H₂O(lane 14).

[0061]FIG. 21B depicts RT-PCR analysis of breast tumor genes in prostatetumors (lane 1, 2), colon tumors (lane 3), lung tumor (lane 4), normalprostate (lane 5), normal colon (lane 6), normal kidney (lane 7), normalliver (lane 8), normal lung (lane 9), normal ovary (lanes 10, 18),normal pancreases (lanes 11, 12), normal skeletal muscle (lane 13),normal skin (lane 14), normal stomach (lane 15), normal testes (lane16), normal small intestine (lane 17), HBL-100 (lane 19), MCF-12A (lane20), breast tumors (lanes 21-23), H₂O (lane 24), and colon tumor (lane25).

[0062]FIG. 22 shows the recognition of a B11Ag1 peptide (referred to asB11-8) by an anti-B11-8 CTL line.

[0063]FIG. 23 shows the recognition of a cell line transduced with theantigen B11Ag1 by the B11-8 specific clone A1.

[0064]FIG. 24 shows recognition of a lung adenocarcinoma line(LT-140-22) and a breast adenocarcinoma line (CAMA-1) by the B11-8specific clone A1.

DETAILED DESCRIPTION OF THE INVENTION

[0065] U.S. patents, U.S. patent application publications, U.S. patentapplications, foreign patents, foreign patent applications andnon-patent publications referred to in this specification and/or listedin the Application Data Sheet, are incorporated herein by reference, intheir entirety.

[0066] The present invention is directed generally to compositions andtheir use in the therapy and diagnosis of cancer, particularly breastcancer. As described further below, illustrative compositions of thepresent invention include, but are not restricted to, polypeptides,particularly immunogenic polypeptides, polynucleotides encoding suchpolypeptides, antibodies and other binding agents, antigen presentingcells (APCs) and immune system cells (e.g., T cells).

[0067] The practice of the present invention will employ, unlessindicated specifically to the contrary, conventional methods ofvirology, immunology, microbiology, molecular biology and recombinantDNA techniques within the skill of the art, many of which are describedbelow for the purpose of illustration. Such techniques are explainedfully in the literature. See, e.g., Sambrook, et al., Molecular Cloning:A Laboratory Manual (2nd Edition, 1989); Maniatis et al., MolecularCloning: A Laboratory Manual (1982); DNA Cloning: A Practical Approach,vol. I & II (D. Glover, ed.); Oligonucleotide Synthesis (N. Gait, ed.,1984); Nucleic Acid Hybridization (B. Hames & S. Higgins, eds., 1985);Transcription and Translation (B. Hames & S. Higgins, eds., 1984);Animal Cell Culture (R. Freshney, ed., 1986); Perbal, A Practical Guideto Molecular Cloning (1984).

[0068] All publications, patents and patent applications cited herein,whether supra or infra, are hereby incorporated by reference in theirentirety.

[0069] As used in this specification and the appended claims, thesingular forms “a,” “an” and “the” include plural references unless thecontent clearly dictates otherwise.

[0070] Polypeptide Compositions

[0071] As used herein, the term “polypeptide” is used in itsconventional meaning, i.e., as a sequence of amino acids. Thepolypeptides are not limited to a specific length of the product; thus,peptides, oligopeptides, and proteins are included within the definitionof polypeptide, and such terms may be used interchangeably herein unlessspecifically indicated otherwise. This term also does not refer to orexclude post-expression modifications of the polypeptide, for example,glycosylations, acetylations, phosphorylations and the like, as well asother modifications known in the art, both naturally occurring andnon-naturally occurring. A polypeptide may be an entire protein, or asubsequence thereof. Particular polypeptides of interest in the contextof this invention are amino acid subsequences comprising epitopes, i.e.,antigenic determinants substantially responsible for the immunogenicproperties of a polypeptide and being capable of evoking an immuneresponse.

[0072] Particularly illustrative polypeptides of the present inventioncomprise those encoded by a polynucleotide sequence set forth in any oneof SEQ ID NO: 1, 3-86, 142-298, 301-303, 307, 313, 314, 316, 317, 323,325, 327-330, 335, 339, and 341-344, or a sequence that hybridizes undermoderately stringent conditions, or, alternatively, under highlystringent conditions, to a polynucleotide sequence set forth in any oneof SEQ ID NO: 1, 3-86, 142-298, 301-303, 307, 313, 314, 316, 317, 323,325, 327-330, 335, 339, and 341-344. Certain other illustrativepolypeptides of the invention comprise amino acid sequences as set forthin any one of SEQ ID NO: 131-140, 299, 300, 304-306, 308-312, 315, 318,324, 326, 331-334, 336, 340, and 345-428.

[0073] The polypeptides of the present invention are sometimes hereinreferred to as breast tumor proteins or breast tumor polypeptides, as anindication that their identification has been based at least in partupon their increased levels of expression in breast tumor samples. Thus,a “breast tumor polypeptide” or “breast tumor protein,” refers generallyto a polypeptide sequence of the present invention, or a polynucleotidesequence encoding such a polypeptide, that is expressed in a substantialproportion of breast tumor samples, for example preferably greater thanabout 20%, more preferably greater than about 30%, and most preferablygreater than about 50% or more of breast tumor samples tested, at alevel that is at least two fold, and preferably at least five fold,greater than the level of expression in normal tissues, as determinedusing a representative assay provided herein. A breast tumor polypeptidesequence of the invention, based upon its increased level of expressionin tumor cells, has particular utility both as a diagnostic marker aswell as a therapeutic target, as further described below.

[0074] In certain preferred embodiments, the polypeptides of theinvention are immunogenic, i.e., they react detectably within animmunoassay (such as an ELISA or T-cell stimulation assay) with antiseraand/or T-cells from a patient with breast cancer. Screening forimmunogenic activity can be performed using techniques well known to theskilled artisan. For example, such screens can be performed usingmethods such as those described in Harlow and Lane, Antibodies: ALaboratory Manual, Cold Spring Harbor Laboratory, 1988. In oneillustrative example, a polypeptide may be immobilized on a solidsupport and contacted with patient sera to allow binding of antibodieswithin the sera to the immobilized polypeptide. Unbound sera may then beremoved and bound antibodies detected using, for example, ¹²⁵I-labeledProtein A.

[0075] As would be recognized by the skilled artisan, immunogenicportions of the polypeptides disclosed herein are also encompassed bythe present invention. An “immunogenic portion,” as used herein, is afragment of an immunogenic polypeptide of the invention that itself isimmunologically reactive (i.e., specifically binds) with the B-cellsand/or T-cell surface antigen receptors that recognize the polypeptide.Immunogenic portions may generally be identified using well knowntechniques, such as those summarized in Paul, Fundamental Immunology,3rd ed., 243-247 (Raven Press, 1993) and references cited therein. Suchtechniques include screening polypeptides for the ability to react withantigen-specific antibodies, antisera and/or T-cell lines or clones. Asused herein, antisera and antibodies are “antigen-specific” if theyspecifically bind to an antigen (i.e., they react with the protein in anELISA or other immunoassay, and do not react detectably with unrelatedproteins). Such antisera and antibodies may be prepared as describedherein, and using well-known techniques.

[0076] In one preferred embodiment, an immunogenic portion of apolypeptide of the present invention is a portion that reacts withantisera and/or T-cells at a level that is not substantially less thanthe reactivity of the full-length polypeptide (e.g., in an ELISA and/orT-cell reactivity assay). Preferably, the level of immunogenic activityof the immunogenic portion is at least about 50%, preferably at leastabout 70% and most preferably greater than about 90% of theimmunogenicity for the full-length polypeptide. In some instances,preferred immunogenic portions will be identified that have a level ofimmunogenic activity greater than that of the corresponding full-lengthpolypeptide, e.g., having greater than about 100% or 150% or moreimmunogenic activity.

[0077] In certain other embodiments, illustrative immunogenic portionsmay include peptides in which an N-terminal leader sequence and/ortransmembrane domain have been deleted. Other illustrative immunogenicportions will contain a small N- and/or C-terminal deletion (e.g., 1-30amino acids, preferably 5-15 amino acids), relative to the matureprotein.

[0078] In another embodiment, a polypeptide composition of the inventionmay also comprise one or more polypeptides that are immunologicallyreactive with T cells and/or antibodies generated against a polypeptideof the invention, particularly a polypeptide having an amino acidsequence disclosed herein, or to an immunogenic fragment or variantthereof.

[0079] In another embodiment of the invention, polypeptides are providedthat comprise one or more polypeptides that are capable of eliciting Tcells and/or antibodies that are immunologically reactive with one ormore polypeptides described herein, or one or more polypeptides encodedby contiguous nucleic acid sequences contained in the polynucleotidesequences disclosed herein, or immunogenic fragments or variantsthereof, or to one or more nucleic acid sequences which hybridize to oneor more of these sequences under conditions of moderate to highstringency.

[0080] The present invention, in another aspect, provides polypeptidefragments comprising at least about 5, 10, 15, 20, 25, 50, or 100contiguous amino acids, or more, including all intermediate lengths, ofa polypeptide compositions set forth herein, such as those set forth inSEQ ID NO: 131-140, 299, 300, 304-306, 308-312, 315, 318, 324, 326,331-334, 336, 340, and 345-428, or those encoded by a polynucleotidesequence set forth in a sequence of SEQ ID NO: 1, 3-86, 142-298,301-303, 307, 313, 314, 316, 317, 323, 325, 327-330, 335, 339, and341-344.

[0081] In another aspect, the present invention provides variants of thepolypeptide compositions described herein. Polypeptide variantsgenerally encompassed by the present invention will typically exhibit atleast about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% or more identity (determined as described below), along itslength, to a polypeptide sequences set forth herein.

[0082] In one preferred embodiment, the polypeptide fragments andvariants provided by the present invention are immunologically reactivewith an antibody and/or T-cell that reacts with a full-lengthpolypeptide specifically set forth herein.

[0083] In another preferred embodiment, the polypeptide fragments andvariants provided by the present invention exhibit a level ofimmunogenic activity of at least about 50%, preferably at least about70%, and most preferably at least about 90% or more of that exhibited bya full-length polypeptide sequence specifically set forth herein.

[0084] A polypeptide “variant,” as the term is used herein, is apolypeptide that typically differs from a polypeptide specificallydisclosed herein in one or more substitutions, deletions, additionsand/or insertions. Such variants may be naturally occurring or may besynthetically generated, for example, by modifying one or more of theabove polypeptide sequences of the invention and evaluating theirimmunogenic activity as described herein and/or using any of a number oftechniques well known in the art.

[0085] For example, certain illustrative variants of the polypeptides ofthe invention include those in which one or more portions, such as anN-terminal leader sequence or transmembrane domain, have been removed.Other illustrative variants include variants in which a small portion(e.g., 1-30 amino acids, preferably 5-15 amino acids) has been removedfrom the N- and/or C-terminal of the mature protein.

[0086] In many~instances, a variant will contain conservativesubstitutions. A “conservative substitution” is one in which an aminoacid is substituted for another amino acid that has similar properties,such that one skilled in the art of peptide chemistry would expect thesecondary structure and hydropathic nature of the polypeptide to besubstantially unchanged. As described above, modifications may be madein the structure of the polynucleotides and polypeptides of the presentinvention and still obtain a functional molecule that encodes a variantor derivative polypeptide with desirable characteristics, e.g., withimmunogenic characteristics. When it is desired to alter the amino acidsequence of a polypeptide to create an equivalent, or even an improved,immunogenic variant or portion of a polypeptide of the invention, oneskilled in the art will typically change one or more of the codons ofthe encoding DNA sequence according to Table 1.

[0087] For example, certain amino acids may be substituted for otheramino acids in a protein structure without appreciable loss ofinteractive binding capacity with structures such as, for example,antigen-binding regions of antibodies or binding sites on substratemolecules. Since it is the interactive capacity and nature of a proteinthat defines that protein's biological functional activity, certainamino acid sequence substitutions can be made in a protein sequence,and, of course, its underlying DNA coding sequence, and neverthelessobtain a protein with like properties. It is thus contemplated thatvarious changes may be made in the peptide sequences of the disclosedcompositions, or corresponding DNA sequences which encode said peptideswithout appreciable loss of their biological utility or activity. TABLE1 Amino Acids Codons Alanine Ala A GCA GCC GCG GCU Cysteine Cys C UGCUGU Aspartic acid Asp D GAG GAU Glutamic acid Glu E GAA GAGPhenylalanine Phe F UUC UUU Glycine Gly G GGA GGC GGG GGU Histidine HisH CAC CAU Isoleucine Ile I AUA AUC AUU Lysine Lys K AAA AAG Leucine LeuL UUA UUG CUA CUC CUG CUU Methionine Met M AUG Asparagine Asn N AAC AAUProline Pro P CCA CCC CCG CCU Glutamine Gln Q CAA CAG Arginine Arg R AGAAGG CGA CGC CGG CGU Serine Ser S AGC AGU UCA UCC UCG UCU Threonine Thr TACA ACC ACG ACU Valine Val V GUA GUC GUG GUU Tryptophan Trp W UGGTyrosine Tyr Y UAC UAU

[0088] In making such changes, the hydropathic index of amino acids maybe considered. The importance of the hydropathic amino acid index inconferring interactive biologic function on a protein is generallyunderstood in the art (Kyte and Doolittle, 1982, incorporated herein byreference). It is accepted that the relative hydropathic character ofthe amino acid contributes to the secondary structure of the resultantprotein, which in turn defines the interaction of the protein with othermolecules, for example, enzymes, substrates, receptors, DNA, antibodies,antigens, and the like. Each amino acid has been assigned a hydropathicindex on the basis of its hydrophobicity and charge characteristics(Kyte and Doolittle, 1982). These values are: isoleucine (+4.5); valine(+4.2); leucine (+3.8); phenylalanine (+2.8); cysteine/cystine (+2.5);methionine (+1.9); alanine (+1.8); glycine (−0.4); threonine (−0.7);serine (−0.8); tryptophan (−0.9); tyrosine (−1.3); proline (−1.6);histidine (−3.2); glutamate (−3.5); glutamine (−3.5); aspartate (−3.5);asparagine (−3.5); lysine (−3.9); and arginine (−4.5).

[0089] It is known in the art that certain amino acids may besubstituted by other amino acids having a similar hydropathic index orscore and still result in a protein with similar biological activity,i.e., still obtain a biological functionally equivalent protein. Inmaking such changes, the substitution of amino acids whose hydropathicindices are within ±2 is preferred, those within ±1 are particularlypreferred, and those within ±0.5 are even more particularly preferred.It is also understood in the art that the substitution of like aminoacids can be made effectively on the basis of hydrophilicity. U.S. Pat.No. 4,554,101 (specifically incorporated herein by reference in itsentirety), states that the greatest local average hydrophilicity of aprotein, as governed by the hydrophilicity of its adjacent amino acids,correlates with a biological property of the protein.

[0090] As detailed in U.S. Pat. No. 4,554,101, the followinghydrophilicity values have been assigned to amino acid residues:arginine (+3.0); lysine (+3.0); aspartate (+3.0±1); glutamate (+3.0±1);serine (+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0);threonine (−0.4); proline (−0.5±1); alanine (−0.5); histidine (−0.5);cysteine (−1.0); methionine (−1.3); valine (−1.5); leucine (−1.8);isoleucine (−1.8); tyrosine (−2.3); phenylalanine (−2.5); tryptophan(−3.4). It is understood that an amino acid can be substituted foranother having a similar hydrophilicity value and still obtain abiologically equivalent, and in particular, an immunologicallyequivalent protein. In such changes, the substitution of amino acidswhose hydrophilicity values are within ±2 is preferred, those within ±1are particularly preferred, and those within ±0.5 are even moreparticularly preferred.

[0091] As outlined above, amino acid substitutions are generallytherefore based on the relative similarity of the amino acid side-chainsubstituents, for example, their hydrophobicity, hydrophilicity, charge,size, and the like. Exemplary substitutions that take various of theforegoing characteristics into consideration are well known to those ofskill in the art and include: arginine and lysine; glutamate andaspartate; serine and threonine; glutamine and asparagine; and valine,leucine and isoleucine.

[0092] In addition, any polynucleotide may be further modified toincrease stability in vivo. Possible modifications include, but are notlimited to, the addition of flanking sequences at the 5′ and/or 3′ ends;the use of phosphorothioate or 2′ O-methyl rather than phosphodiesteraselinkages in the backbone; and/or the inclusion of nontraditional basessuch as inosine, queosine and wybutosine, as well as acetyl- methyl-,thio- and other modified forms of adenine, cytidine, guanine, thymineand uridine.

[0093] Amino acid substitutions may further be made on the basis ofsimilarity in polarity, charge, solubility, hydrophobicity,hydrophilicity and/or the amphipathic nature of the residues. Forexample, negatively charged amino acids include aspartic acid andglutamic acid; positively charged amino acids include lysine andarginine; and amino acids with uncharged polar head groups havingsimilar hydrophilicity values include leucine, isoleucine and valine;glycine and alanine; asparagine and glutamine; and serine, threonine,phenylalanine and tyrosine. Other groups of amino acids that mayrepresent conservative changes include: (1) ala, pro, gly, glu, asp,gin, asn, ser, thr; (2) cys, ser, tyr, thr; (3) val, ile, leu, met, ala,phe; (4) lys, arg, his; and (5) phe, tyr, trp, his. A variant may also,or alternatively, contain nonconservative changes. In a preferredembodiment, variant polypeptides differ from a native sequence bysubstitution, deletion or addition of five amino acids or fewer.Variants may also (or alternatively) be modified by, for example, thedeletion or addition of amino acids that have minimal influence on theimmunogenicity, secondary structure and hydropathic nature of thepolypeptide.

[0094] As noted above, polypeptides may comprise a signal (or leader)sequence at the N-terminal end of the protein, which co-translationallyor post-translationally directs transfer of the protein. The polypeptidemay also be conjugated to a linker or other sequence for ease ofsynthesis, purification or identification of the polypeptide (e.g.,poly-His), or to enhance binding of the polypeptide to a solid support.For example, a polypeptide may be conjugated to an immunoglobulin Fcregion.

[0095] When comparing polypeptide sequences, two sequences are said tobe “identical” if the sequence of amino acids in the two sequences isthe same when aligned for maximum correspondence, as described below.Comparisons between two sequences are typically performed by comparingthe sequences over a comparison window to identify and compare localregions of sequence similarity. A “comparison window” as used herein,refers to a segment of at least about 20 contiguous positions, usually30 to about 75, 40 to about 50, in which a sequence may be compared to areference sequence of the same number of contiguous positions after thetwo sequences are optimally aligned.

[0096] Optimal alignment of sequences for comparison may be conductedusing the Megalign program in the Lasergene suite of bioinformaticssoftware (DNASTAR, Inc., Madison, Wis.), using default parameters. Thisprogram embodies several alignment schemes described in the followingreferences: Dayhoff, M. O. (1978) A model of evolutionary change inproteins—Matrices for detecting distant relationships. In Dayhoff, M. O.(ed.) Atlas of Protein Sequence and Structure, National BiomedicalResearch Foundation, Washington D.C. Vol. 5, Suppl. 3, pp. 345-358; HeinJ. (1990) Unified Approach to Alignment and Phylogenes pp. 626-645Methods in Enzymology vol. 183, Academic Press, Inc., San Diego, Calif.;Higgins, D. G. and Sharp, P. M. (1989) CABIOS 5:151-153; Myers, E. W.and Muller W. (1988) CABIOS 4:11-17; Robinson, E. D. (1971) Comb. Theor11:1051; Saitou, N. Nei, M. (1987) Mol. Biol. Evol. 4:406-425; Sneath,P. H. A. and Sokal, R. R. (1973) Numerical Taxonomy—the Principles andPractice of Numerical Taxonomy, Freeman Press, San Francisco, Calif.;Wilbur, W. J. and Lipman, D. J. (1983) Proc. Natl. Acad., Sci. USA80:726-730.

[0097] Alternatively, optimal alignment of sequences for comparison maybe conducted by the local identity algorithm of Smith and Waterman(1981) Add. APL. Math 2:482, by the identity alignment algorithm ofNeedleman and Wunsch (1970) J. Mol. Biol. 48:443, by the search forsimilarity methods of Pearson and Lipman (1988) Proc. Natl. Acad. Sci.USA 85: 2444, by computerized implementations of these algorithms (GAP,BESTFIT, BLAST, FASTA, and TFASTA in the Wisconsin Genetics SoftwarePackage, Genetics Computer Group (GCG), 575 Science Dr., Madison, Wis.),or by inspection.

[0098] One preferred example of algorithms that are suitable fordetermining percent sequence identity and sequence similarity are theBLAST and BLAST 2.0 algorithms, which are described in Altschul et al.(1977) Nucl. Acids Res. 25:3389-3402 and Altschul et al. (1990) J. Mol.Biol. 215:403-410, respectively. BLAST and BLAST 2.0 can be used, forexample with the parameters described herein, to determine percentsequence identity for the polynucleotides and polypeptides of theinvention. Software for performing BLAST analyses is publicly availablethrough the National Center for Biotechnology Information. For aminoacid sequences, a scoring matrix can be used to calculate the cumulativescore. Extension of the word hits in each direction are halted when: thecumulative alignment score falls off by the quantity X from its maximumachieved value; the cumulative score goes to zero or below, due to theaccumulation of one or more negative-scoring residue alignments; or theend of either sequence is reached. The BLAST algorithm parameters W, Tand X determine the sensitivity and speed of the alignment.

[0099] In one preferred approach, the “percentage of sequence identity”is determined by comparing two optimally aligned sequences over a windowof comparison of at least 20 positions, wherein the portion of thepolypeptide sequence in the comparison window may comprise additions ordeletions (i.e., gaps) of 20 percent or less, usually 5 to 15 percent,or 10 to 12 percent, as compared to the reference sequences (which doesnot comprise additions or deletions) for optimal alignment of the twosequences. The percentage is calculated by determining the number ofpositions at which the identical amino acid residue occurs in bothsequences to yield the number of matched positions, dividing the numberof matched positions by the total number of positions in the referencesequence (i.e., the window size) and multiplying the results by 100 toyield the percentage of sequence identity.

[0100] Within other illustrative embodiments, a polypeptide may be axenogeneic polypeptide that comprises an polypeptide having substantialsequence identity, as described above, to the human polypeptide (alsotermed autologous antigen) which served as a reference polypeptide, butwhich xenogeneic polypeptide is derived from a different, non-humanspecies. One skilled in the art will recognize that “self” antigens areoften poor stimulators of CD8+ and CD4+ T-lymphocyte responses, andtherefore efficient immunotherapeutic strategies directed against tumorpolypeptides require the development of methods to overcome immunetolerance to particular self tumor polypeptides. For example, humansimmunized with prostase protein from a xenogeneic (non human) origin arecapable of mounting an immune response against the counterpart humanprotein, e.g., the human prostase tumor protein present on human tumorcells. Accordingly, the present invention provides methods for purifyingthe xenogeneic form of the tumor proteins set forth herein, such as thepolypeptides set forth in SEQ ID NO: 131-140, 299, 300, 304-306,308-312, 315, 318, 324, 326, 331-334, 336, 340, and 345-428, or thoseencoded by polynucleotide sequences set forth in SEQ ID NO: 1, 3-86,142-298, 301-303, 307, 313, 314, 316, 317, 323, 325, 327-330, 335, 339,and 341-344.

[0101] Therefore, one aspect of the present invention providesxenogeneic variants of the polypeptide compositions described herein.Such xenogeneic variants generally encompassed by the present inventionwill typically exhibit at least about 70%, 75%, 80%, 85%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, or 99% or more identity along theirlengths, to a polypeptide sequences set forth herein.

[0102] More particularly, the invention is directed to mouse, rat,monkey, porcine and other non-human polypeptides which can be used asxenogeneic forms of human polypeptides set forth herein, to induceimmune responses directed against tumor polypeptides of the invention.

[0103] Within other illustrative embodiments, a polypeptide may be afusion polypeptide that comprises multiple polypeptides as describedherein, or that comprises at least one polypeptide as described hereinand an unrelated sequence, such as a known tumor protein. A fusionpartner may, for example, assist in providing T helper epitopes (animmunological fusion partner), preferably T helper epitopes recognizedby humans, or may assist in expressing the protein (an expressionenhancer) at higher yields than the native recombinant protein. Certainpreferred fusion partners are both immunological and expressionenhancing fusion partners. Other fusion partners may be selected so asto increase the solubility of the polypeptide or to enable thepolypeptide to be targeted to desired intracellular compartments. Stillfurther fusion partners include affinity tags, which facilitatepurification of the polypeptide.

[0104] Fusion polypeptides may generally be prepared using standardtechniques, including chemical conjugation. Preferably, a fusionpolypeptide is expressed as a recombinant polypeptide, allowing theproduction of increased levels, relative to a non-fused polypeptide, inan expression system. Briefly, DNA sequences encoding the polypeptidecomponents may be assembled separately, and ligated into an appropriateexpression vector. The 3′ end of the DNA sequence encoding onepolypeptide component is ligated, with or without a peptide linker, tothe 5′ end of a DNA sequence encoding the second polypeptide componentso that the reading frames of the sequences are in phase. This permitstranslation into a single fusion polypeptide that retains the biologicalactivity of both component polypeptides.

[0105] A peptide linker sequence may be employed to separate the firstand second polypeptide components by a distance sufficient to ensurethat each polypeptide folds into its secondary and tertiary structures.Such a peptide linker sequence is incorporated into the fusionpolypeptide using standard techniques well known in the art. Suitablepeptide linker sequences may be chosen based on the following factors:(1) their ability to adopt a flexible extended conformation; (2) theirinability to adopt a secondary structure that could interact withfunctional epitopes on the first and second polypeptides; and (3) thelack of hydrophobic or charged residues that might react with thepolypeptide functional epitopes. Preferred peptide linker sequencescontain Gly, Asn and Ser residues. Other near neutral amino acids, suchas Thr and Ala may also be used in the linker sequence. Amino acidsequences which may be usefully employed as linkers include thosedisclosed in Maratea et al., Gene 40:39-46, 1985; Murphy et al., Proc.Nat. Acad. Sci. USA 83:8258-8262, 1986; U.S. Pat. No. 4,935,233 and U.S.Pat. No. 4,751,180. The linker sequence may generally be from 1 to about50 amino acids in length. Linker sequences are not required when thefirst and second polypeptides have non-essential N-terminal amino acidregions that can be used to separate the functional domains and preventsteric interference.

[0106] The ligated DNA sequences are operably linked to suitabletranscriptional or translational regulatory elements. The regulatoryelements responsible for expression of DNA are located only 5′ to theDNA sequence encoding the first polypeptides. Similarly, stop codonsrequired to end translation and transcription termination signals areonly present 3′ to the DNA sequence encoding the second polypeptide.

[0107] The fusion polypeptide can comprise a polypeptide as describedherein together with an unrelated immunogenic protein, such as animmunogenic protein capable of eliciting a recall response. Examples ofsuch proteins include tetanus, tuberculosis and hepatitis proteins (see,for example, Stoute et al. New Engl. J. Med., 336:86-91, 1997).

[0108] In one preferred embodiment, the immunological fusion partner isderived from a Mycobacterium sp., such as a Mycobacteriumtuberculosis-derived Ra12 fragment. Ra12 compositions and methods fortheir use in enhancing the expression and/or immunogenicity ofheterologous polynucleotide/polypeptide sequences is described in U.S.Patent Application 60/158,585, the disclosure of which is incorporatedherein by reference in its entirety. Briefly, Ra12 refers to apolynucleotide region that is a subsequence of a Mycobacteriumtuberculosis MTB32A nucleic acid. MTB32A is a serine protease of 32 KDmolecular weight encoded by a gene in virulent and avirulent strains ofM. tuberculosis. The nucleotide sequence and amino acid sequence ofMTB32A have been described (for example, U.S. Patent Application60/158,585; see also, Skeiky et al., Infection and Immun. (1999)67:3998-4007, incorporated herein by reference). C-terminal fragments ofthe MTB32A coding sequence express at high levels and remain as asoluble polypeptides throughout the purification process. Moreover, Ra12may enhance the immunogenicity of heterologous immunogenic polypeptideswith which it is fused. One preferred Ra12 fusion polypeptide comprisesa 14 KD C-terminal fragment corresponding to amino acid residues 192 to323 of MTB32A. Other preferred Ra12 polynucleotides generally compriseat least about 15 consecutive nucleotides, at least about 30nucleotides, at least about 60 nucleotides, at least about 100nucleotides, at least about 200 nucleotides, or at least about 300nucleotides that encode a portion of a Ra12 polypeptide. Ra12polynucleotides may comprise a native sequence (i.e., an endogenoussequence that encodes a Ra12 polypeptide or a portion thereof) or maycomprise a variant of such a sequence. Ra12 polynucleotide variants maycontain one or more substitutions, additions, deletions and/orinsertions such that the biological activity of the encoded fusionpolypeptide is not substantially diminished, relative to a fusionpolypeptide comprising a native Ra12 polypeptide. Variants preferablyexhibit at least about 70% identity, more preferably at least about 80%identity and most preferably at least about 90% identity to apolynucleotide sequence that encodes a native Ra12 polypeptide or aportion thereof.

[0109] Within other preferred embodiments, an immunological fusionpartner is derived from protein D, a surface protein of thegram-negative bacterium Haemophilus influenza B (WO 91/18926).Preferably, a protein D derivative comprises approximately the firstthird of the protein (e.g., the first N-terminal 100-110 amino acids),and a protein D derivative may be lipidated. Within certain preferredembodiments, the first 109 residues of a Lipoprotein D fusion partner isincluded on the N-terminus to provide the polypeptide with additionalexogenous T-cell epitopes and to increase the expression level in E.coli (thus functioning as an expression enhancer). The lipid tailensures optimal presentation of the antigen to antigen presenting cells.Other fusion partners include the non-structural protein from influenzaevirus, NS1 (hemaglutinin). Typically, the N-terminal 81 amino acids areused, although different fragments that include T-helper epitopes may beused.

[0110] In another embodiment, the immunological fusion partner is theprotein known as LYTA, or a portion thereof (preferably a C-terminalportion). LYTA is derived from Streptococcus pneumoniae, whichsynthesizes an N-acetyl-L-alanine amidase known as amidase LYTA (encodedby the LytA gene; Gene 43:265-292, 1986). LYTA is an autolysin thatspecifically degrades certain bonds in the peptidoglycan backbone. TheC-terminal domain of the LYTA protein is responsible for the affinity tothe choline or to some choline analogues such as DEAE. This property hasbeen exploited for the development of E. coli C-LYTA expressing plasmidsuseful for expression of fusion proteins. Purification of hybridproteins containing the C-LYTA fragment at the amino terminus has beendescribed (see Biotechnology 10:795-798, 1992). Within a preferredembodiment, a repeat portion of LYTA may be incorporated into a fusionpolypeptide. A repeat portion is found in the C-terminal region startingat residue 178. A particularly preferred repeat portion incorporatesresidues 188-305.

[0111] Yet another illustrative embodiment involves fusion polypeptides,and the polynucleotides encoding them, wherein the fusion partnercomprises a targeting signal capable of directing a polypeptide to theendosomal/lysosomal compartment, as described in U.S. Pat. No.5,633,234. An immunogenic polypeptide of the invention, when fused withthis targeting signal, will associate more efficiently with MHC class IImolecules and thereby provide enhanced in vivo stimulation of CD4⁺T-cells specific for the polypeptide.

[0112] Polypeptides of the invention are prepared using any of a varietyof well known synthetic and/or recombinant techniques, the latter ofwhich are further described below. Polypeptides, portions and othervariants generally less than about 150 amino acids can be generated bysynthetic means, using techniques well known to those of ordinary skillin the art. In one illustrative example, such polypeptides aresynthesized using any of the commercially available solid-phasetechniques, such as the Merrifield solid-phase synthesis method, whereamino acids are sequentially added to a growing amino acid chain. SeeMerrifield, J. Am. Chem. Soc. 85:2149-2146,1963. Equipment for automatedsynthesis of polypeptides is commercially available from suppliers suchas Perkin Elmer/Applied Biosystems Division (Foster City, Calif.), andmay be operated according to the manufacturer's instructions.

[0113] In general, polypeptide compositions (including fusionpolypeptides) of the invention are isolated. An “isolated” polypeptideis one that is removed from its original environment. For example, anaturally-occurring protein or polypeptide is isolated if it isseparated from some or all of the coexisting materials in the naturalsystem. Preferably, such polypeptides are also purified, e.g., are atleast about 90% pure, more preferably at least about 95% pure and mostpreferably at least about 99% pure.

[0114] Polynucleotide Compositions

[0115] The present invention, in other aspects, provides polynucleotidecompositions. The terms “DNA” and “polynucleotide” are used essentiallyinterchangeably herein to refer to a DNA molecule that has been isolatedfree of total genomic DNA of a particular species. “Isolated,” as usedherein, means that a polynucleotide is substantially away from othercoding sequences, and that the DNA molecule does not contain largeportions of unrelated coding DNA, such as large chromosomal fragments orother functional genes or polypeptide coding regions. Of course, thisrefers to the DNA molecule as originally isolated, and does not excludegenes or coding regions later added to the segment by the hand of man.

[0116] As will be understood by those skilled in the art, thepolynucleotide compositions of this invention can include genomicsequences, extra-genomic and plasmid-encoded sequences and smallerengineered gene segments that express, or may be adapted to express,proteins, polypeptides, peptides and the like. Such segments may benaturally isolated, or modified synthetically by the hand of man.

[0117] As will be also recognized by the skilled artisan,polynucleotides of the invention may be single-stranded (coding orantisense) or double-stranded, and may be DNA (genomic, cDNA orsynthetic) or RNA molecules. RNA molecules may include HnRNA molecules,which contain introns and correspond to a DNA molecule in a one-to-onemanner, and mRNA molecules, which do not contain introns. Additionalcoding or non-coding sequences may, but need not, be present within apolynucleotide of the present invention, and a polynucleotide may, butneed not, be linked to other molecules and/or support materials.

[0118] Polynucleotides may comprise a native sequence (i.e., anendogenous sequence that encodes a polypeptide/protein of the inventionor a portion thereof) or may comprise a sequence that encodes a variantor derivative, preferably and immunogenic variant or derivative, of sucha sequence.

[0119] Therefore, according to another aspect of the present invention,polynucleotide compositions are provided that comprise some or all of apolynucleotide sequence set forth in any one of SEQ ID NO: 1, 3-86,142-298, 301-303, 307, 313, 314, 316, 317, 323, 325, 327-330, 335, 339,and 341-344, complements of a polynucleotide sequence set forth in anyone of SEQ ID NO: 1, 3-86, 142-298, 301-303, 307, 313, 314, 316, 317,323, 325, 327-330, 335, 339, and 341-344, and degenerate variants of apolynucleotide sequence set forth in any one of SEQ ID NO: 1, 3-86,142-298, 301-303, 307, 313, 314, 316, 317, 323, 325, 327-330, 335, 339,and 341-344. In certain preferred embodiments, the polynucleotidesequences set forth herein encode immunogenic polypeptides, as describedabove.

[0120] In other related embodiments, the present invention providespolynucleotide variants having substantial identity to the sequencesdisclosed herein in SEQ ID NO: 1, 3-86, 142-298, 301-303, 307, 313, 314,316, 317, 323, 325, 327-330, 335, 339, and 341-344, for example thosecomprising at least 70% sequence identity, preferably at least 75%, 80%,85%, 90%, 95%, 96%, 97%, 98%, or 99% or higher, sequence identitycompared to a polynucleotide sequence of this invention using themethods described herein, (e.g., BLAST analysis using standardparameters, as described below). One skilled in this art will recognizethat these values can be appropriately adjusted to determinecorresponding identity of proteins encoded by two nucleotide sequencesby taking into account codon degeneracy, amino acid similarity, readingframe positioning and the like.

[0121] Typically, polynucleotide variants will contain one or moresubstitutions, additions, deletions and/or insertions, preferably suchthat the immunogenicity of the polypeptide encoded by the variantpolynucleotide is not substantially diminished relative to a polypeptideencoded by a polynucleotide sequence specifically set forth herein). Theterm “variants” should also be understood to encompasses homologousgenes of xenogenic origin.

[0122] In additional embodiments, the present invention providespolynucleotide fragments comprising or consisting of various lengths ofcontiguous stretches of sequence identical to or complementary to one ormore of the sequences disclosed herein. For example, polynucleotides areprovided by this invention that comprise or consist of at least about10, 15, 20, 30, 40, 50, 75, 100, 150, 200, 300, 400, 500 or 1000 or morecontiguous nucleotides of one or more of the sequences disclosed hereinas well as all intermediate lengths there between. It will be readilyunderstood that “intermediate lengths”, in this context, means anylength between the quoted values, such as 16, 17, 18, 19, etc.; 21, 22,23, etc.; 30, 31, 32, etc.; 50, 51, 52, 53, etc.; 100, 101, 102, 103,etc.; 150, 151, 152, 153, etc.; including all integers through 200-500;500-1,000, and the like. A polynucleotide sequence as described here maybe extended at one or both ends by additional nucleotides not found inthe native sequence. This additional sequence may consist of 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleotidesat either end of the disclosed sequence or at both ends of the disclosedsequence.

[0123] In another embodiment of the invention, polynucleotidecompositions are provided that are capable of hybridizing under moderateto high stringency conditions to a polynucleotide sequence providedherein, or a fragment thereof, or a complementary sequence thereof.Hybridization techniques are well known in the art of molecular biology.For purposes of illustration, suitable moderately stringent conditionsfor testing the hybridization of a polynucleotide of this invention withother polynucleotides include prewashing in a solution of 5×SSC, 0.5%SDS, 1.0 mM EDTA (pH 8.0); hybridizing at 50° C.-60° C., 5×SSC,overnight; followed by washing twice at 65° C. for 20 minutes with eachof 2×, 0.5× and 0.2×SSC containing 0.1% SDS. One skilled in the art willunderstand that the stringency of hybridization can be readilymanipulated, such as by altering the salt content of the hybridizationsolution and/or the temperature at which the hybridization is performed.For example, in another embodiment, suitable highly stringenthybridization conditions include those described above, with theexception that the temperature of hybridization is increased, e.g., to60-65° C. or 65-70° C.

[0124] In certain preferred embodiments, the polynucleotides describedabove, e.g., polynucleotide variants, fragments and hybridizingsequences, encode polypeptides that are immunologically cross-reactivewith a polypeptide sequence specifically set forth herein. In otherpreferred embodiments, such polynucleotides encode polypeptides thathave a level of immunogenic activity of at least about 50%, preferablyat least about 70%, and more preferably at least about 90% of that for apolypeptide sequence specifically set forth herein.

[0125] The polynucleotides of the present invention, or fragmentsthereof, regardless of the length of the coding sequence itself, may becombined with other DNA sequences, such as promoters, polyadenylationsignals, additional restriction enzyme sites, multiple cloning sites,other coding segments, and the like, such that their overall length mayvary considerably. It is therefore contemplated that a nucleic acidfragment of almost any length may be employed, with the total lengthpreferably being limited by the ease of preparation and use in theintended recombinant DNA protocol. For example, illustrativepolynucleotide segments with total lengths of about 10,000, about 5000,about 3000, about 2,000, about 1,000, about 500, about 200, about 100,about 50 base pairs in length, and the like, (including all intermediatelengths) are contemplated to be useful in many implementations of thisinvention.

[0126] When comparing polynucleotide sequences, two sequences are saidto be “identical” if the sequence of nucleotides in the two sequences isthe same when aligned for maximum correspondence, as described below.Comparisons between two sequences are typically performed by comparingthe sequences over a comparison window to identify and compare localregions of sequence similarity. A “comparison window” as used herein,refers to a segment of at least about 20 contiguous positions, usually30 to about 75, 40 to about 50, in which a sequence may be compared to areference sequence of the same number of contiguous positions after thetwo sequences are optimally aligned.

[0127] Optimal alignment of sequences for comparison may be conductedusing the Megalign program in the Lasergene suite of bioinformaticssoftware (DNASTAR, Inc., Madison, Wis.), using default parameters. Thisprogram embodies several alignment schemes described in the followingreferences: Dayhoff, M. O. (1978) A model of evolutionary change inproteins—Matrices for detecting distant relationships. In Dayhoff, M. O.(ed.) Atlas of Protein Sequence and Structure, National BiomedicalResearch Foundation, Washington D.C. Vol. 5, Suppl. 3, pp. 345-358; HeinJ. (1990) Unified Approach to Alignment and Phylogenes pp. 626-645Methods in Enzymology vol. 183, Academic Press, Inc., San Diego, Calif.;Higgins, D. G. and Sharp, P. M. (1989) CABIOS 5:151-153; Myers, E. W.and Muller W. (1988) CABIOS 4:11-17; Robinson, E. D. (1971) Comb. Theor11:105; Santou, N. Nes, M. (1987) Mol. Biol. Evol. 4:406-425; Sneath, P.H. A. and Sokal, R. R. (1973) Numerical Taxonomy—the Principles andPractice of Numerical Taxonomy, Freeman Press, San Francisco, Calif.;Wilbur, W. J. and Lipman, D. J. (1983) Proc. Natl. Acad., Sci. USA80:726-730.

[0128] Alternatively, optimal alignment of sequences for comparison maybe conducted by the local identity algorithm of Smith and Waterman(1981) Add. APL. Math 2:482, by the identity alignment algorithm ofNeedleman and Wunsch (1970) J. Mol. Biol. 48:443, by the search forsimilarity methods of Pearson and Lipman (1988) Proc. Natl. Acad. Sci.USA 85: 2444, by computerized implementations of these algorithms (GAP,BESTFIT, BLAST, FASTA, and TFASTA in the Wisconsin Genetics SoftwarePackage, Genetics Computer Group (GCG), 575 Science Dr., Madison, Wis.),or by inspection.

[0129] One preferred example of algorithms that are suitable fordetermining percent sequence identity and sequence similarity are theBLAST and BLAST 2.0 algorithms, which are described in Altschul et al.(1977) Nucl. Acids Res. 25:3389-3402 and Altschul et al. (1990) J. Mol.Biol. 215:403-410, respectively. BLAST and BLAST 2.0 can be used, forexample with the parameters described herein, to determine percentsequence identity for the polynucleotides of the invention. Software forperforming BLAST analyses is publicly available through the NationalCenter for Biotechnology Information. In one illustrative example,cumulative scores can be calculated using, for nucleotide sequences, theparameters M (reward score for a pair of matching residues; always >0)and N (penalty score for mismatching residues; always <0). Extension ofthe word hits in each direction are halted when: the cumulativealignment score falls off by the quantity X from its maximum achievedvalue; the cumulative score goes to zero or below, due to theaccumulation of one or more negative-scoring residue alignments; or theend of either sequence is reached. The BLAST algorithm parameters W, Tand X determine the sensitivity and speed of the alignment. The BLASTNprogram (for nucleotide sequences) uses as defaults a wordlength (W) of11, and expectation (E) of 10, and the BLOSUM62 scoring matrix (seeHenikoff and Henikoff (1989) Proc. Natl. Acad. Sci. USA 89:10915)alignments, (B) of 50, expectation (E) of 10, M=5, N=−4 and a comparisonof both strands.

[0130] Preferably, the “percentage of sequence identity” is determinedby comparing two optimally aligned sequences over a window of comparisonof at least 20 positions, wherein the portion of the polynucleotidesequence in the comparison window may comprise additions or deletions(i.e., gaps) of 20 percent or less, usually 5 to 15 percent, or 10 to 12percent, as compared to the reference sequences (which does not compriseadditions or deletions) for optimal alignment of the two sequences. Thepercentage is calculated by determining the number of positions at whichthe identical nucleic acid bases occurs in both sequences to yield thenumber of matched positions, dividing the number of matched positions bythe total number of positions in the reference sequence (i.e., thewindow size) and multiplying the results by 100 to yield the percentageof sequence identity.

[0131] It will be appreciated by those of ordinary skill in the artthat, as a result of the degeneracy of the genetic code, there are manynucleotide sequences that encode a polypeptide as described herein. Someof these polynucleotides bear minimal homology to the nucleotidesequence of any native gene. Nonetheless, polynucleotides that vary dueto differences in codon usage are specifically contemplated by thepresent invention. Further, alleles of the genes comprising thepolynucleotide sequences provided herein are within the scope of thepresent invention. Alleles are endogenous genes that are altered as aresult of one or more mutations, such as deletions, additions and/orsubstitutions of nucleotides. The resulting mRNA and protein may, butneed not, have an altered structure or function. Alleles may beidentified using standard techniques (such as hybridization,amplification and/or database sequence comparison).

[0132] Therefore, in another embodiment of the invention, a mutagenesisapproach, such as site-specific mutagenesis, is employed for thepreparation of immunogenic variants and/or derivatives of thepolypeptides described herein. By this approach, specific modificationsin a polypeptide sequence can be made through mutagenesis of theunderlying polynucleotides that encode them. These techniques provides astraightforward approach to prepare and test sequence variants, forexample, incorporating one or more of the foregoing considerations, byintroducing one or more nucleotide sequence changes into thepolynucleotide.

[0133] Site-specific mutagenesis allows the production of mutantsthrough the use of specific oligonucleotide sequences which encode theDNA sequence of the desired mutation, as well as a sufficient number ofadjacent nucleotides, to provide a primer sequence of sufficient sizeand sequence complexity to form a stable duplex on both sides of thedeletion junction being traversed. Mutations may be employed in aselected polynucleotide sequence to improve, alter, decrease, modify, orotherwise change the properties of the polynucleotide itself, and/oralter the properties, activity, composition, stability, or primarysequence of the encoded polypeptide.

[0134] In certain embodiments of the present invention, the inventorscontemplate the mutagenesis of the disclosed polynucleotide sequences toalter one or more properties of the encoded polypeptide, such as theimmunogenicity of a polypeptide vaccine. The techniques of site-specificmutagenesis are well-known in the art, and are widely used to createvariants of both polypeptides and polynucleotides. For example,site-specific mutagenesis is often used to alter a specific portion of aDNA molecule. In such embodiments, a primer comprising typically about14 to about 25 nucleotides or so in length is employed, with about 5 toabout 10 residues on both sides of the junction of the sequence beingaltered.

[0135] As will be appreciated by those of skill in the art,site-specific mutagenesis techniques have often employed a phage vectorthat exists in both a single stranded and double stranded form. Typicalvectors useful in site-directed mutagenesis include vectors such as theM13 phage. These phage are readily commercially-available and their useis generally well-known to those skilled in the art. Double-strandedplasmids are also routinely employed in site directed mutagenesis thateliminates the step of transferring the gene of interest from a plasmidto a phage.

[0136] In general, site-directed mutagenesis in accordance herewith isperformed by first obtaining a single-stranded vector or melting apartof two strands of a double-stranded vector that includes within itssequence a DNA sequence that encodes the desired peptide. Anoligonucleotide primer bearing the desired mutated sequence is prepared,generally synthetically. This primer is then annealed with thesingle-stranded vector, and subjected to DNA polymerizing enzymes suchas E. coli polymerase I Klenow fragment, in order to complete thesynthesis of the mutation-bearing strand. Thus, a heteroduplex is formedwherein one strand encodes the original non-mutated sequence and thesecond strand bears the desired mutation. This heteroduplex vector isthen used to transform appropriate cells, such as E. coli cells, andclones are selected which include recombinant vectors bearing themutated sequence arrangement.

[0137] The preparation of sequence variants of the selectedpeptide-encoding DNA segments using site-directed mutagenesis provides ameans of producing potentially useful species and is not meant to belimiting as there are other ways in which sequence variants of peptidesand the DNA sequences encoding them may be obtained. For example,recombinant vectors encoding the desired peptide sequence may be treatedwith mutagenic agents, such as hydroxylamine, to obtain sequencevariants. Specific details regarding these methods and protocols arefound in the teachings of Maloy et al., 1994; Segal, 1976; Prokop andBajpai, 1991; Kuby, 1994; and Maniatis et al., 1982, each incorporatedherein by reference, for that purpose.

[0138] As used herein, the term “oligonucleotide directed mutagenesisprocedure” refers to template-dependent processes and vector-mediatedpropagation which result in an increase in the concentration of aspecific nucleic acid molecule relative to its initial concentration, orin an increase in the concentration of a detectable signal, such asamplification. As used herein, the term “oligonucleotide directedmutagenesis procedure” is intended to refer to a process that involvesthe template-dependent extension of a primer molecule. The term templatedependent process refers to nucleic acid synthesis of an RNA or a DNAmolecule wherein the sequence of the newly synthesized strand of nucleicacid is dictated by the well-known rules of complementary base pairing(see, for example, Watson, 1987). Typically, vector mediatedmethodologies involve the introduction of the nucleic acid fragment intoa DNA or RNA vector, the clonal amplification of the vector, and therecovery of the amplified nucleic acid fragment. Examples of suchmethodologies are provided by U.S. Pat. No. 4,237,224, specificallyincorporated herein by reference in its entirety.

[0139] In another approach for the production of polypeptide variants ofthe present invention, recursive sequence recombination, as described inU.S. Pat. No. 5,837,458, may be employed. In this approach, iterativecycles of recombination and screening or selection are performed to“evolve” individual polynucleotide variants of the invention having, forexample, enhanced immunogenic activity.

[0140] In other embodiments of the present invention, the polynucleotidesequences provided herein can be advantageously used as probes orprimers for nucleic acid hybridization. As such, it is contemplated thatnucleic acid segments that comprise or consist of a sequence region ofat least about a 15 nucleotide long contiguous sequence that has thesame sequence as, or is complementary to, a 15 nucleotide longcontiguous sequence disclosed herein will find particular utility.Longer contiguous identical or complementary sequences, e.g., those ofabout 20, 30, 40, 50, 100, 200, 500, 1000 (including all intermediatelengths) and even up to full length sequences will also be of use incertain embodiments.

[0141] The ability of such nucleic acid probes to specifically hybridizeto a sequence of interest will enable them to be of use in detecting thepresence of complementary sequences in a given sample. However, otheruses are also envisioned, such as the use of the sequence informationfor the preparation of mutant species primers, or primers for use inpreparing other genetic constructions.

[0142] Polynucleotide molecules having sequence regions consisting ofcontiguous nucleotide stretches of 10-14, 15-20, 30, 50, or even of100-200 nucleotides or so (including intermediate lengths as well),identical or complementary to a polynucleotide sequence disclosedherein, are particularly contemplated as hybridization probes for usein, e.g., Southern and Northern blotting. This would allow a geneproduct, or fragment thereof, to be analyzed, both in diverse cell typesand also in various bacterial cells. The total size of fragment, as wellas the size of the complementary stretch(es), will ultimately depend onthe intended use or application of the particular nucleic acid segment.Smaller fragments will generally find use in hybridization embodiments,wherein the length of the contiguous complementary region may be varied,such as between about 15 and about 100 nucleotides, but largercontiguous complementarity stretches may be used, according to thelength complementary sequences one wishes to detect.

[0143] The use of a hybridization probe of about 15-25 nucleotides inlength allows the formation of a duplex molecule that is both stable andselective. Molecules having contiguous complementary sequences overstretches greater than 15 bases in length are generally preferred,though, in order to increase stability and selectivity of the hybrid,and thereby improve the quality and degree of specific hybrid moleculesobtained. One will generally prefer to design nucleic acid moleculeshaving gene-complementary stretches of 15 to 25 contiguous nucleotides,or even longer where desired.

[0144] Hybridization probes may be selected from any portion of any ofthe sequences disclosed herein. All that is required is to review thesequences set forth herein, or to any continuous portion of thesequences, from about 15-25 nucleotides in length up to and includingthe full length sequence, that one wishes to utilize as a probe orprimer. The choice of probe and primer sequences may be governed byvarious factors. For example, one may wish to employ primers fromtowards the termini of the total sequence.

[0145] Small polynucleotide segments or fragments may be readilyprepared by, for example, directly synthesizing the fragment by chemicalmeans, as is commonly practiced using an automated oligonucleotidesynthesizer. Also, fragments may be obtained by application of nucleicacid reproduction technology, such as the PCR™ technology of U.S. Pat.No. 4,683,202 (incorporated herein by reference), by introducingselected sequences into recombinant vectors for recombinant production,and by other recombinant DNA techniques generally known to those ofskill in the art of molecular biology.

[0146] The nucleotide sequences of the invention may be used for theirability to selectively form duplex molecules with complementarystretches of the entire gene or gene fragments of interest. Depending onthe application envisioned, one will typically desire to employ varyingconditions of hybridization to achieve varying degrees of selectivity ofprobe towards target sequence. For applications requiring highselectivity, one will typically desire to employ relatively stringentconditions to form the hybrids, e.g., one will select relatively lowsalt and/or high temperature conditions, such as provided by a saltconcentration of from about 0.02 M to about 0.15 M salt at temperaturesof from about 50° C. to about 70° C. Such selective conditions toleratelittle, if any, mismatch between the probe and the template or targetstrand, and would be particularly suitable for isolating relatedsequences.

[0147] Of course, for some applications, for example, where one desiresto prepare mutants employing a mutant primer strand hybridized to anunderlying template, less stringent (reduced stringency) hybridizationconditions will typically be needed in order to allow formation of theheteroduplex. In these circumstances, one may desire to employ saltconditions such as those of from about 0.15 M to about 0.9 M salt, attemperatures ranging from about 20° C. to about 55° C. Cross-hybridizingspecies can thereby be readily identified as positively hybridizingsignals with respect to control hybridizations. In any case, it isgenerally appreciated that conditions can be rendered more stringent bythe addition of increasing amounts of formamide, which serves todestabilize the hybrid duplex in the same manner as increasedtemperature. Thus, hybridization conditions can be readily manipulated,and thus will generally be a method of choice depending on the desiredresults.

[0148] According to another embodiment of the present invention,polynucleotide compositions comprising antisense oligonucleotides areprovided. Antisense oligonucleotides have been demonstrated to beeffective and targeted inhibitors of protein synthesis, and,consequently, provide a therapeutic approach by which a disease can betreated by inhibiting the synthesis of proteins that contribute to thedisease. The efficacy of antisense oligonucleotides for inhibitingprotein synthesis is well established. For example, the synthesis ofpolygalactauronase and the muscarine type 2 acetylcholine receptor areinhibited by antisense oligonucleotides directed to their respectivemRNA sequences (U.S. Pat. No. 5,739,119 and U.S. Pat. No. 5,759,829).Further, examples of antisense inhibition have been demonstrated withthe nuclear protein cyclin, the multiple drug resistance gene (MDG1),ICAM-1, E-selectin, STK-1, striatal GABA_(A) receptor and human EGF(Jaskulski et al., Science. Jun. 10, 1988; 240(4858):1544-6;Vasanthakumar and Ahmed, Cancer Commun. 1989;1(4):225-32; Peris et al.,Brain Res Mol Brain Res. Jun. 15, 1998;57(2):310-20; U.S. Pat. No.5,801,154; U.S. Pat. No. 5,789,573; U.S. Pat. No. 5,718,709 and U.S.Pat. No. 5,610,288). Antisense constructs have also been described thatinhibit and can be used to treat a variety of abnormal cellularproliferations, e.g., cancer (U.S. Pat. No. 5,747,470; U.S. Pat. No.5,591,317 and U.S. Pat. No. 5,783,683).

[0149] Therefore, in certain embodiments, the present invention providesoligonucleotide sequences that comprise all, or a portion of, anysequence that is capable of specifically binding to polynucleotidesequence described herein, or a complement thereof. In one embodiment,the antisense oligonucleotides comprise DNA or derivatives thereof. Inanother embodiment, the oligonucleotides comprise RNA or derivativesthereof. In a third embodiment, the oligonucleotides are modified DNAscomprising a phosphorothioated modified backbone. In a fourthembodiment, the oligonucleotide sequences comprise peptide nucleic acidsor derivatives thereof. In each case, preferred compositions comprise asequence region that is complementary, and more preferablysubstantially-complementary, and even more preferably, completelycomplementary to one or more portions of polynucleotides disclosedherein. Selection of antisense compositions specific for a given genesequence is based upon analysis of the chosen target sequence anddetermination of secondary structure, T_(m), binding energy, andrelative stability. Antisense compositions may be selected based upontheir relative inability to form dimers, hairpins, or other secondarystructures that would reduce or prohibit specific binding to the targetmRNA in a host cell. Highly preferred target regions of the mRNA, arethose which are at or near the AUG translation initiation codon, andthose sequences which are substantially complementary to 5′ regions ofthe mRNA. These secondary structure analyses and target site selectionconsiderations can be performed, for example, using v.4 of the OLIGOprimer analysis software and/or the BLASTN 2.0.5 algorithm software(Altschul et al., Nucleic Acids Res. 1997, 25(17):3389-402).

[0150] The use of an antisense delivery method employing a short peptidevector, termed MPG (27 residues), is also contemplated. The MPG peptidecontains a hydrophobic domain derived from the fusion sequence of HIVgp41 and a hydrophilic domain from the nuclear localization sequence ofSV40 T-antigen (Morris et al., Nucleic Acids Res. Jul. 15,1997;25(14):2730-6). It has been demonstrated that several molecules ofthe MPG peptide coat the antisense oligonucleotides and can be deliveredinto cultured mammalian cells in less than 1 hour with relatively highefficiency (90%). Further, the interaction with MPG strongly increasesboth the stability of the oligonucleotide to nuclease and the ability tocross the plasma membrane.

[0151] According to another embodiment of the invention, thepolynucleotide compositions described herein are used in the design andpreparation of ribozyme molecules for inhibiting expression of the tumorpolypeptides and proteins of the present invention in tumor cells.Ribozymes are RNA-protein complexes that cleave nucleic acids in asite-specific fashion. Ribozymes have specific catalytic domains thatpossess endonuclease activity (Kim and Cech, Proc Natl Acad Sci USA.December 1987;84(24):8788-92; Forster and Symons, Cell. Apr. 24,1987;49(2):211-20). For example, a large number of ribozymes acceleratephosphoester transfer reactions with a high degree of specificity, oftencleaving only one of several phosphoesters in an oligonucleotidesubstrate (Cech et al., Cell. December 1981;27(3 Pt 2):487-96; Micheland Westhof, J Mol Biol. Dec. 5, 1990;216(3):585-610; Reinhold-Hurek andShub, Nature. May 14, 1992; 357(6374):173-6). This specificity has beenattributed to the requirement that the substrate bind via specificbase-pairing interactions to the internal guide sequence (“IGS”) of theribozyme prior to chemical reaction.

[0152] Six basic varieties of naturally-occurring enzymatic RNAs areknown presently. Each can catalyze the hydrolysis of RNA phosphodiesterbonds in trans (and thus can cleave other RNA molecules) underphysiological conditions. In general, enzymatic nucleic acids act byfirst binding to a target RNA. Such binding occurs through the targetbinding portion of a enzymatic nucleic acid which is held in closeproximity to an enzymatic portion of the molecule that acts to cleavethe target RNA. Thus, the enzymatic nucleic acid first recognizes andthen binds a target RNA through complementary base-pairing, and oncebound to the correct site, acts enzymatically to cut the target RNA.Strategic cleavage of such a target RNA will destroy its ability todirect synthesis of an encoded protein. After an enzymatic nucleic acidhas bound and cleaved its RNA target, it is released from that RNA tosearch for another target and can repeatedly bind and cleave newtargets.

[0153] The enzymatic nature of a ribozyme is advantageous over manytechnologies, such as antisense technology (where a nucleic acidmolecule simply binds to a nucleic acid target to block its translation)since the concentration of ribozyme necessary to affect a therapeutictreatment is lower than that of an antisense oligonucleotide. Thisadvantage reflects the ability of the ribozyme to act enzymatically.Thus, a single ribozyme molecule is able to cleave many molecules oftarget RNA. In addition, the ribozyme is a highly specific inhibitor,with the specificity of inhibition depending not only on the basepairing mechanism of binding to the target RNA, but also on themechanism of target RNA cleavage. Single mismatches, orbase-substitutions, near the site of cleavage can completely eliminatecatalytic activity of a ribozyme. Similar mismatches in antisensemolecules do not prevent their action (Woolf et al., Proc Natl Acad SciUSA. Aug. 15, 1992;89(16):7305-9). Thus, the specificity of action of aribozyme is greater than that of an antisense oligonucleotide bindingthe same RNA site.

[0154] The enzymatic nucleic acid molecule may be formed in ahammerhead, hairpin, a hepatitis δ virus, group I intron or RNaseP RNA(in association with an RNA guide sequence) or Neurospora VS RNA motif.Examples of hammerhead motifs are described by Rossi et al., NucleicAcids Res. Sep. 11, 1992;20(17):4559-65. Examples of hairpin motifs aredescribed by Hampel et al. (Eur. Pat. Appl. Publ. No. EP 0360257),Hampel and Tritz, Biochemistry Jun. 13, 1989;28(12):4929-33; Hampel etal., Nucleic Acids Res. Jan. 25, 1990;18(2):299-304 and U.S. Pat. No.5,631,359. An example of the hepatitis δ virus motif is described byPerrotta and Been, Biochemistry. Dec. 1, 1992; 31(47):11843-52; anexample of the RNaseP motif is described by Guerrier-Takada et al.,Cell. December 1983;35(3 Pt 2):849-57; Neurospora VS RNA ribozyme motifis described by Collins (Saville and Collins, Cell. May 18,1990;61(4):685-96; Saville and Collins, Proc Natl Acad Sci USA. Oct. 1,1991;88(19):8826-30; Collins and Olive, Biochemistry. Mar. 23,1993;32(11):2795-9); and an example of the Group I intron is describedin (U.S. Pat. No. 4,987,071). All that is important in an enzymaticnucleic acid molecule of this invention is that it has a specificsubstrate binding site which is complementary to one or more of thetarget gene RNA regions, and that it have nucleotide sequences within orsurrounding that substrate binding site which impart an RNA cleavingactivity to the molecule. Thus the ribozyme constructs need not belimited to specific motifs mentioned herein.

[0155] Ribozymes may be designed as described in Int. Pat. Appl. Publ.No. WO 93/23569 and Int. Pat. Appl. Publ. No. WO 94/02595, eachspecifically incorporated herein by reference) and synthesized to betested in vitro and in vivo, as described. Such ribozymes can also beoptimized for delivery. While specific examples are provided, those inthe art will recognize that equivalent RNA targets in other species canbe utilized when necessary.

[0156] Ribozyme activity can be optimized by altering the length of theribozyme binding arms, or chemically synthesizing ribozymes withmodifications that prevent their degradation by serum ribonucleases (seee.g., Int. Pat. Appl. Publ. No. WO 92/07065; lnt. Pat. Appl. Publ. No.WO 93/15187; Int. Pat. Appl. Publ. No. WO 91/03162; Eur. Pat. Appl.Publ. No. 92110298.4; U.S. Pat. No. 5,334,711; and lnt. Pat. Appl. Publ.No. WO 94/13688, which describe various chemical modifications that canbe made to the sugar moieties of enzymatic RNA molecules), modificationswhich enhance their efficacy in cells, and removal of stem II bases toshorten RNA synthesis times and reduce chemical requirements.

[0157] Sullivan et al. (Int. Pat. Appl. Publ. No. WO 94/02595) describesthe general methods for delivery of enzymatic RNA molecules. Ribozymesmay be administered to cells by a variety of methods known to thosefamiliar to the art, including, but not restricted to, encapsulation inliposomes, by iontophoresis, or by incorporation into other vehicles,such as hydrogels, cyclodextrins, biodegradable nanocapsules, andbioadhesive microspheres. For some indications, ribozymes may bedirectly delivered ex vivo to cells or tissues with or without theaforementioned vehicles. Alternatively, the RNA/vehicle combination maybe locally delivered by direct inhalation, by direct injection or by useof a catheter, infusion pump or stent. Other routes of delivery include,but are not limited to, intravascular, intramuscular, subcutaneous orjoint injection, aerosol inhalation, oral (tablet or pill form),topical, systemic, ocular, intraperitoneal and/or intrathecal delivery.More detailed descriptions of ribozyme delivery and administration areprovided in Int. Pat. Appl. Publ. No. WO 94/02595 and Int. Pat. Appl.Publ. No. WO 93/23569, each specifically incorporated herein byreference.

[0158] Another means of accumulating high concentrations of aribozyme(s) within cells is to incorporate the ribozyme-encodingsequences into a DNA expression vector. Transcription of the ribozymesequences are driven from a promoter for eukaryotic RNA polymerase I(pol I ), RNA polymerase II (pol II ), or RNA polymerase III (pol III ).Transcripts from pol II or pol III promoters will be expressed at highlevels in all cells; the levels of a given pol II promoter in a givencell type will depend on the nature of the gene regulatory sequences(enhancers, silencers, etc.) present nearby. Prokaryotic RNA polymerasepromoters may also be used, providing that the prokaryotic RNApolymerase enzyme is expressed in the appropriate cells Ribozymesexpressed from such promoters have been shown to function in mammaliancells. Such transcription units can be incorporated into a variety ofvectors for introduction into mammalian cells, including but notrestricted to, plasmid DNA vectors, viral DNA vectors (such asadenovirus or adeno-associated vectors), or viral RNA vectors (such asretroviral, semliki forest virus, sindbis virus vectors).

[0159] In another embodiment of the invention, peptide nucleic acids(PNAs) compositions are provided. PNA is a DNA mimic in which thenucleobases are attached to a pseudopeptide backbone (Good and Nielsen,Antisense Nucleic Acid Drug Dev. 1997 7(4) 431-37). PNA is able to beutilized in a number methods that traditionally have used RNA or DNA.Often PNA sequences perform better in techniques than the correspondingRNA or DNA sequences and have utilities that are not inherent to RNA orDNA. A review of PNA including methods of making, characteristics of,and methods of using, is provided by Corey (Trends Biotechnol June1997;15(6):224-9). As such, in certain embodiments, one may prepare PNAsequences that are complementary to one or more portions of the ACE mRNAsequence, and such PNA compositions may be used to regulate, alter,decrease, or reduce the translation of ACE-specific mRNA, and therebyalter the level of ACE activity in a host cell to which such PNAcompositions have been administered.

[0160] PNAs have 2-aminoethyl-glycine linkages replacing the normalphosphodiester backbone of DNA (Nielsen et al., Science Dec. 6,1991;254(5037):1497-500; Hanvey et al., Science. Nov. 27,1992;258(5087):1481-5; Hyrup and Nielsen, Bioorg Med Chem. January1996;4(1):5-23). This chemistry has three important consequences:firstly, in contrast to DNA or phosphorothioate oligonucleotides, PNAsare neutral molecules; secondly, PNAs are achiral, which avoids the needto develop a stereoselective synthesis; and thirdly, PNA synthesis usesstandard Boc or Fmoc protocols for solid-phase peptide synthesis,although other methods, including a modified Merrifield method, havebeen used.

[0161] PNA monomers or ready-made oligomers are commercially availablefrom PerSeptive Biosystems (Framingham, Mass.). PNA syntheses by eitherBoc or Fmoc protocols are straightforward using manual or automatedprotocols (Norton et al., Bioorg Med Chem. April 1995;3(4):437-45). Themanual protocol lends itself to the production of chemically modifiedPNAs or the simultaneous synthesis of families of closely related PNAs.

[0162] As with peptide synthesis, the success of a particular PNAsynthesis will depend on the properties of the chosen sequence. Forexample, while in theory PNAs can incorporate any combination ofnucleotide bases, the presence of adjacent purines can lead to deletionsof one or more residues in the product. In expectation of thisdifficulty, it is suggested that, in producing PNAs with adjacentpurines, one should repeat the coupling of residues likely to be addedinefficiently. This should be followed by the purification of PNAs byreverse-phase high-pressure liquid chromatography, providing yields andpurity of product similar to those observed during the synthesis ofpeptides.

[0163] Modifications of PNAs for a given application may be accomplishedby coupling amino acids during solid-phase synthesis or by attachingcompounds that contain a carboxylic acid group to the exposed N-terminalamine. Alternatively, PNAs can be modified after synthesis by couplingto an introduced lysine or cysteine. The ease with which PNAs can bemodified facilitates optimization for better solubility or for specificfunctional requirements. Once synthesized, the identity of PNAs andtheir derivatives can be confirmed by mass spectrometry. Several studieshave made and utilized modifications of PNAs (for example, Norton etal., Bioorg Med Chem. April 1995;3(4):437-45; Petersen et al., J PeptSci. May-June 1995;1(3):175-83; Orum et al., Biotechniques. September1995;19(3):472-80; Footer et al., Biochemistry. Aug. 20,1996;35(33):10673-9; Griffith et al., Nucleic Acids Res. Aug. 11,1995;23(15):3003-8; Pardridge et al., Proc Natl Acad Sci USA. Jun. 6,1995;92(12):5592-6; Boffa et al., Proc Natl Acad Sci USA. Mar. 14,1995;92(6):1901-5; Gambacorti-Passerini et al., Blood. Aug. 15,1996;88(4):1411-7; Armitage et al., Proc Natl Acad Sci USA. Nov. 11,1997;94(23):12320-5; Seeger et al., Biotechniques. September1997;23(3):512-7). U.S. Pat. No. 5,700,922 discusses PNA-DNA-PNAchimeric molecules and their uses in diagnostics, modulating protein inorganisms, and treatment of conditions susceptible to therapeutics.

[0164] Methods of characterizing the antisense binding properties ofPNAs are discussed in Rose (Anal Chem. Dec. 15, 1993;65(24):3545-9) andJensen et al. (Biochemistry. Apr. 22, 1997;36(16):5072-7). Rose usescapillary gel electrophoresis to determine binding of PNAs to theircomplementary oligonucleotide, measuring the relative binding kineticsand stoichiometry. Similar types of measurements were made by Jensen etal. using BIAcore™ technology.

[0165] Other applications of PNAs that have been described and will beapparent to the skilled artisan include use in DNA strand invasion,antisense inhibition, mutational analysis, enhancers of transcription,nucleic acid purification, isolation of transcriptionally active genes,blocking of transcription factor binding, genome cleavage, biosensors,in situ hybridization, and the like.

[0166] Polynucleotide Identification, Characterization and Expression

[0167] Polynucleotides compositions of the present invention may beidentified, prepared and/or manipulated using any of a variety of wellestablished techniques (see generally, Sambrook et al., MolecularCloning: A Laboratory Manual, Cold Spring Harbor Laboratories, ColdSpring Harbor, N.Y., 1989, and other like references). For example, apolynucleotide may be identified, as described in more detail below, byscreening a microarray of cDNAs for tumor-associated expression (i.e.,expression that is at least two fold greater in a tumor than in normaltissue, as determined using a representative assay provided herein).Such screens may be performed, for example, using the microarraytechnology of Affymetrix, Inc. (Santa Clara, Calif.) according to themanufacturer's instructions (and essentially as described by Schena etal., Proc. Natl. Acad. Sci. USA 93:10614-10619, 1996 and Heller et al.,Proc. Natl. Acad. Sci. USA 94:2150-2155, 1997). Alternatively,polynucleotides may be amplified from cDNA prepared from cellsexpressing the proteins described herein, such as tumor cells.

[0168] Many template dependent processes are available to amplify atarget sequences of interest present in a sample. One of the best knownamplification methods is the polymerase chain reaction (PCR™) which isdescribed in detail in U.S. Pat. Nos. 4,683,195, 4,683,202 and4,800,159, each of which is incorporated herein by reference in itsentirety. Briefly, in PCR™, two primer sequences are prepared which arecomplementary to regions on opposite complementary strands of the targetsequence. An excess of deoxynucleoside triphosphates is added to areaction mixture along with a DNA polymerase (e.g., Taq polymerase). Ifthe target sequence is present in a sample, the primers will bind to thetarget and the polymerase will cause the primers to be extended alongthe target sequence by adding on nucleotides. By raising and loweringthe temperature of the reaction mixture, the extended primers willdissociate from the target to form reaction products, excess primerswill bind to the target and to the reaction product and the process isrepeated. Preferably reverse transcription and PCR™ amplificationprocedure may be performed in order to quantify the amount of mRNAamplified. Polymerase chain reaction methodologies are well known in theart.

[0169] Any of a number of other template dependent processes, many ofwhich are variations of the PCR™ amplification technique, are readilyknown and available in the art. Illustratively, some such methodsinclude the ligase chain reaction (referred to as LCR), described, forexample, in Eur. Pat. Appl. Publ. No. 320,308 and U.S. Pat. No.4,883,750; Qbeta Replicase, described in PCT Intl. Pat. Appl. Publ. No.PCT/US87/00880; Strand Displacement Amplification (SDA) and Repair ChainReaction (RCR). Still other amplification methods are described in GreatBritain Pat. Appl. No. 2 202 328, and in PCT Intl. Pat. Appl. Publ. No.PCT/US89/01025. Other nucleic acid amplification procedures includetranscription-based amplification systems (TAS) (PCT Intl. Pat. Appl.Publ. No. WO 88/10315), including nucleic acid sequence basedamplification (NASBA) and 3SR. Eur. Pat. Appl. Publ. No. 329,822describes a nucleic acid amplification process involving cyclicallysynthesizing single-stranded RNA (“ssRNA”), ssDNA, and double-strandedDNA (dsDNA). PCT Intl. Pat. Appl. Publ. No. WO 89/06700 describes anucleic acid sequence amplification scheme based on the hybridization ofa promoter/primer sequence to a target single-stranded DNA (“ssDNA”)followed by transcription of many RNA copies of the sequence. Otheramplification methods such as “RACE” (Frohman, 1990), and “one-sidedPCR” (Ohara, 1989) are also well-known to those of skill in the art.

[0170] An amplified portion of a polynucleotide of the present inventionmay be used to isolate a full length gene from a suitable library (e.g.,a tumor cDNA library) using well known techniques. Within suchtechniques, a library (cDNA or genomic) is screened using one or morepolynucleotide probes or primers suitable for amplification. Preferably,a library is size-selected to include larger molecules. Random primedlibraries may also be preferred for identifying 5′ and upstream regionsof genes. Genomic libraries are preferred for obtaining introns andextending 5′ sequences.

[0171] For hybridization techniques, a partial sequence may be labeled(e.g., by nick-translation or end-labeling with ³²P) using well knowntechniques. A bacterial or bacteriophage library is then generallyscreened by hybridizing filters containing denatured bacterial colonies(or lawns containing phage plaques) with the labeled probe (see Sambrooket al., Molecular Cloning: A Laboratory Manual, Cold Spring HarborLaboratories, Cold Spring Harbor, N.Y., 1989). Hybridizing colonies orplaques are selected and expanded, and the DNA is isolated for furtheranalysis. cDNA clones may be analyzed to determine the amount ofadditional sequence by, for example, PCR using a primer from the partialsequence and a primer from the vector. Restriction maps and partialsequences may be generated to identify one or more overlapping clones.The complete sequence may then be determined using standard techniques,which may involve generating a series of deletion clones. The resultingoverlapping sequences can then assembled into a single contiguoussequence. A full length cDNA molecule can be generated by ligatingsuitable fragments, using well known techniques.

[0172] Alternatively, amplification techniques, such as those describedabove, can be useful for obtaining a full length coding sequence from apartial cDNA sequence. One such amplification technique is inverse PCR(see Triglia et al., Nucl. Acids Res. 16:8186,1988), which usesrestriction enzymes to generate a fragment in the known region of thegene. The fragment is then circularized by intramolecular ligation andused as a template for PCR with divergent primers derived from the knownregion. Within an alternative approach, sequences adjacent to a partialsequence may be retrieved by amplification with a primer to a linkersequence and a primer specific to a known region. The amplifiedsequences are typically subjected to a second round of amplificationwith the same linker primer and a second primer specific to the knownregion. A variation on this procedure, which employs two primers thatinitiate extension in opposite directions from the known sequence, isdescribed in WO 96/38591. Another such technique is known as “rapidamplification of cDNA ends” or RACE. This technique involves the use ofan internal primer and an external primer, which hybridizes to a polyAregion or vector sequence, to identify sequences that are 5′ and 3′ of aknown sequence. Additional techniques include capture PCR (Lagerstrom etal., PCR Methods Applic. 1:111-19, 1991) and walking PCR (Parker et al.,Nucl. Acids. Res. 19:3055-60, 1991). Other methods employingamplification may also be employed to obtain a full length cDNAsequence.

[0173] In certain instances, it is possible to obtain a full length cDNAsequence by analysis of sequences provided in an expressed sequence tag(EST) database, such as that available from GenBank. Searches foroverlapping ESTs may generally be performed using well known programs(e.g., NCBI BLAST searches), and such ESTs may be used to generate acontiguous full length sequence. Full length DNA sequences may also beobtained by analysis of genomic fragments.

[0174] In other embodiments of the invention, polynucleotide sequencesor fragments thereof which encode polypeptides of the invention, orfusion proteins or functional equivalents thereof, may be used inrecombinant DNA molecules to direct expression of a polypeptide inappropriate host cells. Due to the inherent degeneracy of the geneticcode, other DNA sequences that encode substantially the same or afunctionally equivalent amino acid sequence may be produced and thesesequences may be used to clone and express a given polypeptide.

[0175] As will be understood by those of skill in the art, it may beadvantageous in some instances to produce polypeptide-encodingnucleotide sequences possessing non-naturally occurring codons. Forexample, codons preferred by a particular prokaryotic or eukaryotic hostcan be selected to increase the rate of protein expression or to producea recombinant RNA transcript having desirable properties, such as ahalf-life which is longer than that of a transcript generated from thenaturally occurring sequence.

[0176] Moreover, the polynucleotide sequences of the present inventioncan be engineered using methods generally known in the art in order toalter polypeptide encoding sequences for a variety of reasons, includingbut not limited to, alterations which modify the cloning, processing,and/or expression of the gene product. For example, DNA shuffling byrandom fragmentation and PCR reassembly of gene fragments and syntheticoligonucleotides may be used to engineer the nucleotide sequences. Inaddition, site-directed mutagenesis may be used to insert newrestriction sites, alter glycosylation patterns, change codonpreference, produce splice variants, or introduce mutations, and soforth.

[0177] In another embodiment of the invention, natural, modified, orrecombinant nucleic acid sequences may be ligated to a heterologoussequence to encode a fusion protein. For example, to screen peptidelibraries for inhibitors of polypeptide activity, it may be useful toencode a chimeric protein that can be recognized by a commerciallyavailable antibody. A fusion protein may also be engineered to contain acleavage site located between the polypeptide-encoding sequence and theheterologous protein sequence, so that the polypeptide may be cleavedand purified away from the heterologous moiety.

[0178] Sequences encoding a desired polypeptide may be synthesized, inwhole or in part, using chemical methods well known in the art (seeCaruthers, M. H. et al. (1980) Nucl. Acids Res. Symp. Ser. 215-223,Horn, T. et al. (1980) Nucl. Acids Res. Symp. Ser. 225-232).Alternatively, the protein itself may be produced using chemical methodsto synthesize the amino acid sequence of a polypeptide, or a portionthereof. For example, peptide synthesis can be performed using varioussolid-phase techniques (Roberge, J. Y. et al. (1995) Science269:202-204) and automated synthesis may be achieved, for example, usingthe ABI 431A Peptide Synthesizer (Perkin Elmer, Palo Alto, Calif.).

[0179] A newly synthesized peptide may be substantially purified bypreparative high performance liquid chromatography (e.g., Creighton, T.(1983) Proteins, Structures and Molecular Principles, W H Freeman andCo., New York, N.Y.) or other comparable techniques available in theart. The composition of the synthetic peptides may be confirmed by aminoacid analysis or sequencing (e.g., the Edman degradation procedure).Additionally, the amino acid sequence of a polypeptide, or any partthereof, may be altered during direct synthesis and/or combined usingchemical methods with sequences from other proteins, or any partthereof, to produce a variant polypeptide.

[0180] In order to express a desired polypeptide, the nucleotidesequences encoding the polypeptide, or functional equivalents, may beinserted into appropriate expression vector, i.e., a vector whichcontains the necessary elements for the transcription and translation ofthe inserted coding sequence. Methods which are well known to thoseskilled in the art may be used to construct expression vectorscontaining sequences encoding a polypeptide of interest and appropriatetranscriptional and translational control elements. These methodsinclude in vitro recombinant DNA techniques, synthetic techniques, andin vivo genetic recombination. Such techniques are described, forexample, in Sambrook, J. et al. (1989) Molecular Cloning, A LaboratoryManual, Cold Spring Harbor Press, Plainview, N.Y., and Ausubel, F. M. etal. (1989) Current Protocols in Molecular Biology, John Wiley & Sons,New York. N.Y.

[0181] A variety of expression vector/host systems may be utilized tocontain and express polynucleotide sequences. These include, but are notlimited to, microorganisms such as bacteria transformed with recombinantbacteriophage, plasmid, or cosmid DNA expression vectors; yeasttransformed with yeast expression vectors; insect cell systems infectedwith virus expression vectors (e.g., baculovirus); plant cell systemstransformed with virus expression vectors (e.g., cauliflower mosaicvirus, CaMV; tobacco mosaic virus, TMV) or with bacterial expressionvectors (e.g., Ti or pBR322 plasmids); or animal cell systems.

[0182] The “control elements” or “regulatory sequences” present in anexpression vector are those non-translated regions of thevector—enhancers, promoters, 5′ and 3′ untranslated regions—whichinteract with host cellular proteins to carry out transcription andtranslation. Such elements may vary in their strength and specificity.Depending on the vector system and host utilized, any number of suitabletranscription and translation elements, including constitutive andinducible promoters, may be used. For example, when cloning in bacterialsystems, inducible promoters such as the hybrid lacZ promoter of thepBLUESCRIPT phagemid (Stratagene, La Jolla, Calif.) or pSPORT1 plasmid(Gibco BRL, Gaithersburg, Md.) and the like may be used. In mammaliancell systems, promoters from mammalian genes or from mammalian virusesare generally preferred. If it is necessary to generate a cell line thatcontains multiple copies of the sequence encoding a polypeptide, vectorsbased on SV40 or EBV may be advantageously used with an appropriateselectable marker.

[0183] In bacterial systems, any of a number of expression vectors maybe selected depending upon the use intended for the expressedpolypeptide. For example, when large quantities are needed, for examplefor the induction of antibodies, vectors which direct high levelexpression of fusion proteins that are readily purified may be used.Such vectors include, but are not limited to, the multifunctional E.coli cloning and expression vectors such as pBLUESCRIPT (Stratagene), inwhich the sequence encoding the polypeptide of interest may be ligatedinto the vector in frame with sequences for the amino-terminal Met andthe subsequent 7 residues of .beta.-galactosidase so that a hybridprotein is produced; pIN vectors (Van Heeke, G. and S. M. Schuster(1989) J. Biol. Chem. 264:5503-5509); and the like. pGEX Vectors(Promega, Madison, Wis.) may also be used to express foreignpolypeptides as fusion proteins with glutathione S-transferase (GST). Ingeneral, such fusion proteins are soluble and can easily be purifiedfrom lysed cells by adsorption to glutathione-agarose beads followed byelution in the presence of free glutathione. Proteins made in suchsystems may be designed to include heparin, thrombin, or factor XAprotease cleavage sites so that the cloned polypeptide of interest canbe released from the GST moiety at will.

[0184] In the yeast, Saccharomyces cerevisiae, a number of vectorscontaining constitutive or inducible promoters such as alpha factor,alcohol oxidase, and PGH may be used. For reviews,,see Ausubel et al.(supra) and Grant et al. (1987) Methods Enzymol. 153:516-544.

[0185] In cases where plant expression vectors are used, the expressionof sequences encoding polypeptides may be driven by any of a number ofpromoters. For example, viral promoters such as the 35S and 19Spromoters of CaMV may be used alone or in combination with the omegaleader sequence from TMV (Takamatsu, N. (1987) EMBO J. 6:307-311.Alternatively, plant promoters such as the small subunit of RUBISCO orheat shock promoters may be used (Coruzzi, G. et al. (1984) EMBO J.3:1671-1680; Broglie, R. et al. (1984) Science 224:838-843; and Winter,J. et al. (1991) Results Probl. Cell Differ. 17:85-105). Theseconstructs can be introduced into plant cells by direct DNAtransformation or pathogen-mediated transfection. Such techniques aredescribed in a number of generally available reviews (see, for example,Hobbs, S. or Murry, L. E. in McGraw Hill Yearbook of Science andTechnology (1992) McGraw Hill, New York, N.Y.; pp. 191-196).

[0186] An insect system may also be used to express a polypeptide ofinterest. For example, in one such system, Autographa californicanuclear polyhedrosis virus (AcNPV) is used as a vector to expressforeign genes in Spodoptera frugiperda cells or in Trichoplusia larvae.The sequences encoding the polypeptide may be cloned into anon-essential region of the virus, such as the polyhedrin gene, andplaced under control of the polyhedrin promoter. Successful insertion ofthe polypeptide-encoding sequence will render the polyhedrin geneinactive and produce recombinant virus lacking coat protein. Therecombinant viruses may then be used to infect, for example, S.frugiperda cells or Trichoplusia larvae in which the polypeptide ofinterest may be expressed (Engelhard, E. K. et al. (1994) Proc. Natl.Acad. Sci. 91 :3224-3227).

[0187] In mammalian host cells, a number of viral-based expressionsystems are generally available. For example, in cases where anadenovirus is used as an expression vector, sequences encoding apolypeptide of interest may be ligated into an adenovirustranscription/translation complex consisting of the late promoter andtripartite leader sequence. Insertion in a non-essential E1 or E3 regionof the viral genome may be used to obtain a viable virus which iscapable of expressing the polypeptide in infected host cells (Logan, J.and Shenk, T. (1984) Proc. Natl. Acad. Sci. 81:3655-3659). In addition,transcription enhancers, such as the Rous sarcoma virus (RSV) enhancer,may be used to increase expression in mammalian host cells.

[0188] Specific initiation signals may also be used to achieve moreefficient translation of sequences encoding a polypeptide of interest.Such signals include the ATG initiation codon and adjacent sequences. Incases where sequences encoding the polypeptide, its initiation codon,and upstream sequences are inserted into the appropriate expressionvector, no additional transcriptional or translational control signalsmay be needed. However, in cases where only coding sequence, or aportion thereof, is inserted, exogenous translational control signalsincluding the ATG initiation codon should be provided. Furthermore, theinitiation codon should be in the correct reading frame to ensuretranslation of the entire insert. Exogenous translational elements andinitiation codons may be of various origins, both natural and synthetic.The efficiency of expression may be enhanced by the inclusion ofenhancers which are appropriate for the particular cell system which isused, such as those described in the literature (Scharf, D. et al.(1994) Results Probl. Cell Differ. 20:125-162).

[0189] In addition, a host cell strain may be chosen for its ability tomodulate the expression of the inserted sequences or to process theexpressed protein in the desired fashion. Such modifications of thepolypeptide include, but are not limited to, acetylation, carboxylation,glycosylation, phosphorylation, lipidation, and acylation.Post-translational processing which cleaves a “prepro” form of theprotein may also be used to facilitate correct insertion, folding and/orfunction. Different host cells such as CHO, COS, HeLa, MDCK, HEK293, andWI38, which have specific cellular machinery and characteristicmechanisms for such post-translational activities, may be chosen toensure the correct modification and processing of the foreign protein.

[0190] For long-term, high-yield production of recombinant proteins,stable expression is generally preferred. For example, cell lines whichstably express a polynucleotide of interest may be transformed usingexpression vectors which may contain viral origins of replication and/orendogenous expression elements and a selectable marker gene on the sameor on a separate vector. Following the introduction of the vector, cellsmay be allowed to grow for 1-2 days in an enriched media before they areswitched to selective media. The purpose of the selectable marker is toconfer resistance to selection, and its presence allows growth andrecovery of cells which successfully express the introduced sequences.Resistant clones of stably transformed cells may be proliferated usingtissue culture techniques appropriate to the cell type.

[0191] Any number of selection systems may be used to recovertransformed cell lines. These include, but are not limited to, theherpes simplex virus thymidine kinase (Wigler, M. et al. (1977) Cell11:223-32) and adenine phosphoribosyltransferase (Lowy, I. et al. (1990)Cell 22:817-23) genes which can be employed in tk.sup.- oraprt.sup.-cells, respectively. Also, antimetabolite, antibiotic orherbicide resistance can be used as the basis for selection; forexample, dhfr which confers resistance to methotrexate (Wigler, M. etal. (1980) Proc. Natl. Acad. Sci. 77:3567-70); npt, which confersresistance to the aminoglycosides, neomycin and G-418 (Colbere-Garapin,F. et al (1981) J. Mol. Biol. 150:1-14); and als or pat, which conferresistance to chlorsulfuron and phosphinotricin acetyltransferase,respectively (Murry, supra). Additional selectable genes have beendescribed, for example, trpB, which allows cells to utilize indole inplace of tryptophan, or hisD, which allows cells to utilize histinol inplace of histidine (Hartman, S. C. and R. C. Mulligan (1988) Proc. Natl.Acad. Sci. 85:8047-51). The use of visible markers has gained popularitywith such markers as anthocyanins, beta-glucuronidase and its substrateGUS, and luciferase and its substrate luciferin, being widely used notonly to identify transformants, but also to quantify the amount oftransient or stable protein expression attributable to a specific vectorsystem (Rhodes, C. A. et al. (1995) Methods Mol. Biol. 55:121-131).

[0192] Although the presence/absence of marker gene expression suggeststhat the gene of interest is also present, its presence and expressionmay need to be confirmed. For example, if the sequence encoding apolypeptide is inserted within a marker gene sequence, recombinant cellscontaining sequences can be identified by the absence of marker genefunction. Alternatively, a marker gene can be placed in tandem with apolypeptide-encoding sequence under the control of a single promoter.Expression of the marker gene in response to induction or selectionusually indicates expression of the tandem gene as well.

[0193] Alternatively, host cells that contain and express a desiredpolynucleotide sequence may be identified by a variety of proceduresknown to those of skill in the art. These procedures include, but arenot limited to, DNA-DNA or DNA-RNA hybridizations and protein bioassayor immunoassay techniques which include, for example, membrane,solution, or chip based technologies for the detection and/orquantification of nucleic acid or protein.

[0194] A variety of protocols for detecting and measuring the expressionof polynucleotide-encoded products, using either polyclonal ormonoclonal antibodies specific for the product are known in the art.Examples include enzyme-linked immunosorbent assay (ELISA),radioimmunoassay (RIA), and fluorescence activated cell sorting (FACS).A two-site, monoclonal-based immunoassay utilizing monoclonal antibodiesreactive to two non-interfering epitopes on a given polypeptide may bepreferred for some applications, but a competitive binding assay mayalso be employed. These and other assays are described, among otherplaces, in Hampton, R. et al. (1990; Serological Methods, a LaboratoryManual, APS Press, St Paul. Minn.) and Maddox, D. E. et al. (1983; J.Exp. Med. 158:1211-1216).

[0195] A wide variety of labels and conjugation techniques are known bythose skilled in the art and may be used in various nucleic acid andamino acid assays. Means for producing labeled hybridization or PCRprobes for detecting sequences related to polynucleotides includeoligolabeling, nick translation, end-labeling or PCR amplification usinga labeled nucleotide. Alternatively, the sequences, or any portionsthereof may be cloned into a vector for the production of an mRNA probe.Such vectors are known in the art, are commercially available, and maybe used to synthesize RNA probes in vitro by addition of an appropriateRNA polymerase such as T7, T3, or SP6 and labeled nucleotides. Theseprocedures may be conducted using a variety of commercially availablekits. Suitable reporter molecules or labels, which may be used includeradionuclides, enzymes, fluorescent, chemiluminescent, or chromogenicagents as well as substrates, cofactors, inhibitors, magnetic particles,and the like.

[0196] Host cells transformed with a polynucleotide sequence of interestmay be cultured under conditions suitable for the expression andrecovery of the protein from cell culture. The protein produced by arecombinant cell may be secreted or contained intracellularly dependingon the sequence and/or the vector used. As will be understood by thoseof skill in the art, expression vectors containing polynucleotides ofthe invention may be designed to contain signal sequences which directsecretion of the encoded polypeptide through a prokaryotic or eukaryoticcell membrane. Other recombinant constructions may be used to joinsequences encoding a polypeptide of interest to nucleotide sequenceencoding a polypeptide domain which will facilitate purification ofsoluble proteins. Such purification facilitating domains include, butare not limited to, metal chelating peptides such ashistidine-tryptophan modules that allow purification on immobilizedmetals, protein A domains that allow purification on immobilizedimmunoglobulin, and the domain utilized in the FLAGS extension/affinitypurification system (Immunex Corp., Seattle, Wash.). The inclusion ofcleavable linker sequences such as those specific for Factor XA orenterokinase (Invitrogen, San Diego, Calif.) between the purificationdomain and the encoded polypeptide may be used to facilitatepurification. One such expression vector provides for expression of afusion protein containing a polypeptide of interest and a nucleic acidencoding 6 histidine residues preceding a thioredoxin or an enterokinasecleavage site. The histidine residues facilitate purification on IMIAC(immobilized metal ion affinity chromatography) as described in Porath,J. et al. (1992, Prot. Exp. Purif. 3:263-281) while the enterokinasecleavage site provides a means for purifying the desired polypeptidefrom the fusion protein. A discussion of vectors which contain fusionproteins is provided in Kroll, D. J. et al. (1993; DNA Cell Biol.12:441-453).

[0197] In addition to recombinant production methods, polypeptides ofthe invention, and fragments thereof, may be produced by direct peptidesynthesis using solid-phase techniques (Merrifield J. (1963) J. Am.Chem. Soc. 85:2149-2154). Protein synthesis may be performed usingmanual techniques or by automation. Automated synthesis may be achieved,for example, using Applied Biosystems 431A Peptide Synthesizer (PerkinElmer). Alternatively, various fragments may be chemically synthesizedseparately and combined using chemical methods to produce the fulllength molecule.

[0198] Antibidy Compositions, Fragments Thereof and Other Bonding Agents

[0199] According to another aspect, the present invention furtherprovides binding agents, such as antibodies and antigen-bindingfragments thereof, that exhibit immunological binding to a tumorpolypeptide disclosed herein, or to a portion, variant or derivativethereof. An antibody, or antigen-binding fragment thereof, is said to“specifically bind,” “immunogically bind,” and/or is “immunologicallyreactive” to a polypeptide of the invention if it reacts at a detectablelevel (within, for example, an ELISA assay) with the polypeptide, anddoes not react detectably with unrelated polypeptides under similarconditions.

[0200] Immunological binding, as used in this context, generally refersto the non-covalent interactions of the type which occur between animmunoglobulin molecule and an antigen for which the immunoglobulin isspecific. The strength, or affinity of immunological bindinginteractions can be expressed in terms of the dissociation constant(K_(d)) of the interaction, wherein a smaller K_(d) represents a greateraffinity. Immunological binding properties of selected polypeptides canbe quantified using methods well known in the art. One such methodentails measuring the rates of antigen-binding site/antigen complexformation and dissociation, wherein those rates depend on theconcentrations of the complex partners, the affinity of the interaction,and on geometric parameters that equally influence the rate in bothdirections. Thus, both the “on rate constant” (K_(on)) and the “off rateconstant” (K_(off)) can be determined by calculation of theconcentrations and the actual rates of association and dissociation. Theratio of K_(off)/K_(on) enables cancellation of all parameters notrelated to affinity, and is thus equal to the dissociation constantK_(d). See, generally, Davies et al. (1990) Annual Rev. Biochem.59:439-473.

[0201] An “antigen-binding site,” or “binding portion” of an antibodyrefers to the part of the immunoglobulin molecule that participates inantigen binding. The antigen binding site is formed by amino acidresidues of the N-terminal variable (“V”) regions of the heavy (“H”) andlight (“L”) chains. Three highly divergent stretches within the Vregions of the heavy and light chains are referred to as “hypervariableregions” which are interposed between more conserved flanking stretchesknown as “framework regions,” or “FRs”. Thus the term “FR” refers toamino acid sequences which are naturally found between and adjacent tohypervariable regions in immunoglobulins. In an antibody molecule, thethree hypervariable regions of a light chain and the three hypervariableregions of a heavy chain are disposed relative to each other in threedimensional space to form an antigen-binding surface. Theantigen-binding surface is complementary to the three-dimensionalsurface of a bound antigen, and the three hypervariable regions of eachof the heavy and light chains are referred to as“complementarity-determining regions,” or “CDRs.”

[0202] Binding agents may be further capable of differentiating betweenpatients with and without a cancer, such as breast cancer, using therepresentative assays provided herein. For example, antibodies or otherbinding agents that bind to a tumor protein will preferably generate asignal indicating the presence of a cancer in at least about 20% ofpatients with the disease, more preferably at least about 30% ofpatients. Alternatively, or in addition, the antibody will generate anegative signal indicating the absence of the disease in at least about90% of individuals without the cancer. To determine whether a bindingagent satisfies this requirement, biological samples (e.g., blood, sera,sputum, urine and/or tumor biopsies) from patients with and without acancer (as determined using standard clinical tests) may be assayed asdescribed herein for the presence of polypeptides that bind to thebinding agent. Preferably, a statistically significant number of sampleswith and without the disease will be assayed. Each binding agent shouldsatisfy the above criteria; however, those of ordinary skill in the artwill recognize that binding agents may be used in combination to improvesensitivity.

[0203] Any agent that satisfies the above requirements may be a bindingagent. For example, a binding agent may be a ribosome, with or without apeptide component, an RNA molecule or a polypeptide. In a preferredembodiment, a binding agent is an antibody or an antigen-bindingfragment thereof. Antibodies may be prepared by any of a variety oftechniques known to those of ordinary skill in the art. See, e.g.,Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring HarborLaboratory, 1988. In general, antibodies can be produced by cell culturetechniques, including the generation of monoclonal antibodies asdescribed herein, or via transfection of antibody genes into suitablebacterial or mammalian cell hosts, in order to allow for the productionof recombinant antibodies. In one technique, an immunogen comprising thepolypeptide is initially injected into any of a wide variety of mammals(e.g., mice, rats, rabbits, sheep or goats). In this step, thepolypeptides of this invention may serve as the immunogen withoutmodification. Alternatively, particularly for relatively shortpolypeptides, a superior immune response may be elicited if thepolypeptide is joined to a carrier protein, such as bovine serum albuminor keyhole limpet hemocyanin. The immunogen is injected into the animalhost, preferably according to a predetermined schedule incorporating oneor more booster immunizations, and the animals are bled periodically.Polyclonal antibodies specific for the polypeptide may then be purifiedfrom such antisera by, for example, affinity chromatography using thepolypeptide coupled to a suitable solid support.

[0204] Monoclonal antibodies specific for an antigenic polypeptide ofinterest may be prepared, for example, using the technique of Kohler andMilstein, Eur. J. Immunol. 6:511-519, 1976, and improvements thereto.Briefly, these methods involve the preparation of immortal cell linescapable of producing antibodies having the desired specificity (i.e.,reactivity with the polypeptide of interest). Such cell lines may beproduced, for example, from spleen cells obtained from an animalimmunized as described above. The spleen cells are then immortalized by,for example, fusion with a myeloma cell fusion partner, preferably onethat is syngeneic with the immunized animal. A variety of fusiontechniques may be employed. For example, the spleen cells and myelomacells may be combined with a nonionic detergent for a few minutes andthen plated at low density on a selective medium that supports thegrowth of hybrid cells, but not myeloma cells. A preferred selectiontechnique uses HAT (hypoxanthine, aminopterin, thymidine) selection.After a sufficient time, usually about 1 to 2 weeks, colonies of hybridsare observed. Single colonies are selected and their culturesupernatants tested for binding activity against the polypeptide.Hybridomas having high reactivity and specificity are preferred.

[0205] Monoclonal antibodies may be isolated from the supernatants ofgrowing hybridoma colonies. In addition, various techniques may beemployed to enhance the yield, such as injection of the hybridoma cellline into the peritoneal cavity of a suitable vertebrate host, such as amouse. Monoclonal antibodies may then be harvested from the ascitesfluid or the blood. Contaminants may be removed from the antibodies byconventional techniques, such as chromatography, gel filtration,precipitation, and extraction. The polypeptides of this invention may beused in the purification process in, for example, an affinitychromatography step.

[0206] A number of therapeutically useful molecules are known in the artwhich comprise antigen-binding sites that are capable of exhibitingimmunological binding properties of an antibody molecule. Theproteolytic enzyme papain preferentially cleaves IgG molecules to yieldseveral fragments, two of which (the “F(ab)” fragments) each comprise acovalent heterodimer that includes an intact antigen-binding site. Theenzyme pepsin is able to cleave IgG molecules to provide severalfragments, including the “F(ab′)₂” fragment which comprises bothantigen-binding sites. An “Fv” fragment can be produced by preferentialproteolytic cleavage of an IgM, and on rare occasions IgG or IgAimmunoglobulin molecule. Fv fragments are, however, more commonlyderived using recombinant techniques known in the art. The Fv fragmentincludes a non-covalent V_(H)::V_(L) heterodimer including anantigen-binding site which retains much of the antigen recognition andbinding capabilities of the native antibody molecule. Inbar et al.(1972) Proc. Nat. Acad. Sci. USA 69:2659-2662; Hochman et al. (1976)Biochem 15:2706-2710; and Ehrlich et al. (1980) Biochem 19:4091-4096.

[0207] A single chain Fv (“sFv”) polypeptide is a covalently linkedV_(H)::V_(L) heterodimer which is expressed from a gene fusion includingV_(H)- and V_(L)-encoding genes linked by a peptide-encoding linker.Huston et al. (1988) Proc. Nat. Acad. Sci. USA 85(16):5879-5883. Anumber of methods have been described to discern chemical structures forconverting the naturally aggregated—but chemically separated—light andheavy polypeptide chains from an antibody V region into an sFv moleculewhich will fold into a three dimensional structure substantially similarto the structure of an antigen-binding site. See, e.g., U.S. Pat. Nos.5,091,513 and 5,132,405, to Huston et al.; and U.S. Pat. No. 4,946,778,to Ladner et al.

[0208] Each of the above-described molecules includes a heavy chain anda light chain CDR set, respectively interposed between a heavy chain anda light chain FR set which provide support to the CDRS and define thespatial relationship of the CDRs relative to each other. As used herein,the term “CDR set” refers to the three hypervariable regions of a heavyor light chain V region. Proceeding from the N-terminus of a heavy orlight chain, these regions are denoted as “CDR1,” “CDR2,” and “CDR3”respectively. An antigen-binding site, therefore, includes six CDRs,comprising the CDR set from each of a heavy and a light chain V region.A polypeptide comprising a single CDR, (e.g., a CDR1, CDR2 or CDR3) isreferred to herein as a “molecular recognition unit.” Crystallographicanalysis of a number of antigen-antibody complexes has demonstrated thatthe amino acid residues of CDRs form extensive contact with boundantigen, wherein the most extensive antigen contact is with the heavychain CDR3. Thus, the molecular recognition units are primarilyresponsible for the specificity of an antigen-binding site.

[0209] As used herein, the term “FR set” refers to the four flankingamino acid sequences which frame the CDRs of a CDR set of a heavy orlight chain V region. Some FR residues may contact bound antigen;however, FRs are primarily responsible for folding the V region into theantigen-binding site, particularly the FR residues directly adjacent tothe CDRS. Within FRs, certain amino residues and certain structuralfeatures are very highly conserved. In this regard, all V regionsequences contain an internal disulfide loop of around 90 amino acidresidues. When the V regions fold into a binding-site, the CDRs aredisplayed as projecting loop motifs which form an antigen-bindingsurface. It is generally recognized that there are conserved structuralregions of FRs which influence the folded shape of the CDR loops intocertain “canonical” structures—regardless of the precise CDR amino acidsequence. Further, certain FR residues are known to participate innon-covalent interdomain contacts which stabilize the interaction of theantibody heavy and light chains.

[0210] A number of “humanized” antibody molecules comprising anantigen-binding site derived from a non-human immunoglobulin have beendescribed, including chimeric antibodies having rodent V regions andtheir associated CDRs fused to human constant domains (Winter et al.(1991) Nature 349:293-299; Lobuglio et al. (1989) Proc. Nat. Acad. Sci.USA 86:4220-4224; Shaw et al. (1987) J Immunol. 138:4534-4538; and Brownet al. (1987) Cancer Res. 47:3577-3583), rodent CDRs grafted into ahuman supporting FR prior to fusion with an appropriate human antibodyconstant domain (Riechmann et al. (1988) Nature 332:323-327; Verhoeyenet al. (1988) Science 239:1534-1536; and Jones et al. (1986) Nature321:522-525), and rodent CDRs supported by recombinantly veneered rodentFRs (European Patent Publication No. 519,596, published Dec. 23, 1992).These “humanized” molecules are designed to minimize unwantedimmunological response toward rodent antihuman antibody molecules whichlimits the duration and effectiveness of therapeutic applications ofthose moieties in human recipients.

[0211] As used herein, the terms “veneered FRs” and “recombinantlyveneered FRs” refer to the selective replacement of FR residues from,e.g., a rodent heavy or light chain V region, with human FR residues inorder to provide a xenogeneic molecule comprising an antigen-bindingsite which retains substantially all of the native FR polypeptidefolding structure. Veneering techniques are based on the understandingthat the ligand binding characteristics of an antigen-binding site aredetermined primarily by the structure and relative disposition of theheavy and light chain CDR sets within the antigen-binding surface.Davies et al. (1990) Ann. Rev. Biochem. 59:439-473. Thus, antigenbinding specificity can be preserved in a humanized antibody onlywherein the CDR structures, their interaction with each other, and theirinteraction with the rest of the V region domains are carefullymaintained. By using veneering techniques, exterior (e.g.,solvent-accessible) FR residues which are readily encountered by theimmune system are selectively replaced with human residues to provide ahybrid molecule that comprises either a weakly immunogenic, orsubstantially non-immunogenic veneered surface.

[0212] The process of veneering makes use of the available sequence datafor human antibody variable domains compiled by Kabat et al., inSequences of Proteins of Immunological Interest, 4th ed., (U.S. Dept. ofHealth and Human Services, U.S. Government Printing Office, 1987),updates to the Kabat database, and other accessible U.S. and foreigndatabases (both nucleic acid and protein). Solvent accessibilities of Vregion amino acids can be deduced from the known three-dimensionalstructure for human and murine antibody fragments. There are two generalsteps in veneering a murine antigen-binding site. Initially, the FRs ofthe variable domains of an antibody molecule of interest are comparedwith corresponding FR sequences of human variable domains obtained fromthe above-identified sources. The most homologous human V regions arethen compared residue by residue to corresponding murine amino acids.The residues in the murine FR which differ from the human counterpartare replaced by the residues present in the human moiety usingrecombinant techniques well known in the art. Residue switching is onlycarried out with moieties which are at least partially exposed (solventaccessible), and care is exercised in the replacement of amino acidresidues which may have a significant effect on the tertiary structureof V region domains, such as proline, glycine and charged amino acids.

[0213] In this manner, the resultant “veneered” murine antigen-bindingsites are thus designed to retain the murine CDR residues, the residuessubstantially adjacent to the CDRs, the residues identified as buried ormostly buried (solvent inaccessible), the residues believed toparticipate in non-covalent (e.g., electrostatic and hydrophobic)contacts between heavy and light chain domains, and the residues fromconserved structural regions of the FRs which are believed to influencethe “canonical” tertiary structures of the CDR loops. These designcriteria are then used to prepare recombinant nucleotide sequences whichcombine the CDRs of both the heavy and light chain of a murineantigen-binding site into human-appearing FRs that can be used totransfect mammalian cells for the expression of recombinant humanantibodies which exhibit the antigen specificity of the murine antibodymolecule.

[0214] In another embodiment of the invention, monoclonal antibodies ofthe present invention may be coupled to one or more therapeutic agents.Suitable 5 agents in this regard include radionuclides, differentiationinducers, drugs, toxins, and derivatives thereof. Preferredradionuclides include ⁹⁰Y, ¹²³I, ¹²⁵I, ¹³¹I, ¹⁸⁶Re, ¹⁸⁸Re, ²¹¹At, and²¹²Bi. Preferred drugs include methotrexate, and pyrimidine and purineanalogs. Preferred differentiation inducers include phorbol esters andbutyric acid. Preferred toxins include ricin, abrin, diptheria toxin,cholera toxin, gelonin, Pseudomonas exotoxin, Shigella toxin, andpokeweed antiviral protein.

[0215] A therapeutic agent may be coupled (e.g., covalently bonded) to asuitable monoclonal antibody either directly or indirectly (e.g., via alinker group). A direct reaction between an agent and an antibody ispossible when each possesses a substituent capable of reacting with theother. For example, a nucleophilic group, such as an amino or sulfhydrylgroup, on one may be capable of reacting with a carbonyl-containinggroup, such as an anhydride or an acid halide, or with an alkyl groupcontaining a good leaving group (e.g., a halide) on the other.

[0216] Alternatively, it may be desirable to couple a therapeutic agentand an antibody via a linker group. A linker group can function as aspacer to distance an antibody from an agent in order to avoidinterference with binding capabilities. A linker group can also serve toincrease the chemical reactivity of a substituent on an agent or anantibody, and thus increase the coupling efficiency. An increase inchemical reactivity may also facilitate the use of agents, or functionalgroups on agents, which otherwise would not be possible.

[0217] It will be evident to those skilled in the art that a variety ofbifunctional or polyfunctional reagents, both homo- andhetero-functional (such as those described in the catalog of the PierceChemical Co., Rockford, Ill.), may be employed as the linker group.Coupling may be effected, for example, through amino groups, carboxylgroups, sulfhydryl groups or oxidized carbohydrate residues. There arenumerous references describing such methodology, e.g., U.S. Pat. No.4,671,958, to Rodwell et al.

[0218] Where a therapeutic agent is more potent when free from theantibody portion of the immunoconjugates of the present invention, itmay be desirable to use a linker group which is cleavable during or uponinternalization into a cell. A number of different cleavable linkergroups have been described. The mechanisms for the intracellular releaseof an agent from these linker groups include cleavage by reduction of adisulfide bond (e.g., U.S. Pat. No. 4,489,710, to Spitler), byirradiation of a photolabile bond (e.g., U.S. Pat. No. 4,625,014, toSenter et al.), by hydrolysis of derivatized amino acid side chains(e.g., U.S. Pat. No. 4,638,045, to Kohn et al.), by serumcomplement-mediated hydrolysis (e.g., U.S. Pat. No. 4,671,958, toRodwell et al.), and acid-catalyzed hydrolysis (e.g., U.S. Pat. No.4,569,789, to Blattler et al.).

[0219] It may be desirable to couple more than one agent to an antibody.In one embodiment, multiple molecules of an agent are coupled to oneantibody molecule. In another embodiment, more than one type of agentmay be coupled to one antibody. Regardless of the particular embodiment,immunoconjugates with more than one agent may be prepared in a varietyof ways. For example, more than one agent may be coupled directly to anantibody molecule, or linkers that provide multiple sites for attachmentcan be used. Alternatively, a carrier can be used.

[0220] A carrier may bear the agents in a variety of ways, includingcovalent bonding either directly or via a linker group. Suitablecarriers include proteins such as albumins (e.g., U.S. Pat. No.4,507,234, to Kato et al.), peptides and polysaccharides such asaminodextran (e.g., U.S. Pat. No. 4,699,784, to Shih et al.). A carriermay also bear an agent by noncovalent bonding or by encapsulation, suchas within a liposome vesicle (e.g., U.S. Pat. Nos. 4,429,008 and4,873,088). Carriers specific for radionuclide agents includeradiohalogenated small molecules and chelating compounds. For example,U.S. Pat. No. 4,735,792 discloses representative radiohalogenated smallmolecules and their synthesis. A radionuclide chelate may be formed fromchelating compounds that include those containing nitrogen and sulfuratoms as the donor atoms for binding the metal, or metal oxide,radionuclide. For example, U.S. Pat. No. 4,673,562, to Davison et al.discloses representative chelating compounds and their synthesis.

[0221] T Cell Compositions

[0222] The present invention, in another aspect, provides T cellsspecific for a tumor polypeptide disclosed herein, or for a variant orderivative thereof. Such cells may generally be prepared in vitro or exvivo, using standard procedures. For example, T cells may be isolatedfrom bone marrow, peripheral blood, or a fraction of bone marrow orperipheral blood of a patient, using a commercially available cellseparation system, such as the Isolex™ System, available from NexellTherapeutics, Inc. (Irvine, Calif.; see also U.S. Pat. No. 5,240,856;U.S. Pat. No. 5,215,926; WO 89/06280; WO 91/16116 and WO 92/07243).Alternatively, T cells may be derived from related or unrelated humans,non-human mammals, cell lines or cultures.

[0223] T cells may be stimulated with a polypeptide, polynucleotideencoding a polypeptide and/or an antigen presenting cell (APC) thatexpresses such a polypeptide. Such stimulation is performed underconditions and for a time sufficient to permit the generation of T cellsthat are specific for the polypeptide of interest. Preferably, a tumorpolypeptide or polynucleotide of the invention is present within adelivery vehicle, such as a microsphere, to facilitate the generation ofspecific T cells.

[0224] T cells are considered to be specific for a polypeptide of thepresent invention if the T cells specifically proliferate, secretecytokines or kill target cells coated with the polypeptide or expressinga gene encoding the polypeptide. T cell specificity may be evaluatedusing any of a variety of standard techniques. For example, within achromium release assay or proliferation assay, a stimulation index ofmore than two fold increase in lysis and/or proliferation, compared tonegative controls, indicates T cell specificity. Such assays may beperformed, for example, as described in Chen et al., Cancer Res.54:1065-1070, 1994. Alternatively, detection of the proliferation of Tcells may be accomplished by a variety of known techniques. For example,T cell proliferation can be detected by measuring an increased rate ofDNA synthesis (e.g., by pulse-labeling cultures of T cells withtritiated thymidine and measuring the amount of tritiated thymidineincorporated into DNA). Contact with a tumor polypeptide (100 ng/ml-100μg/ml, preferably 200 ng/ml-25 μg/ml) for 3-7 days will typically resultin at least a two fold increase in proliferation of the T cells. Contactas described above for 2-3 hours should result in activation of the Tcells, as measured using standard cytokine assays in which a two foldincrease in the level of cytokine release (e.g., TNF or IFN-γ) isindicative of T cell activation (see Coligan et al., Current Protocolsin Immunology, vol. 1, Wiley lnterscience (Greene 1998)). T cells thathave been activated in response to a tumor polypeptide, polynucleotideor polypeptide-expressing APC may be CD4⁺ and/or CD8⁺. Tumorpolypeptide-specific T cells may be expanded using standard techniques.Within preferred embodiments, the T cells are derived from a patient, arelated donor or an unrelated donor, and are administered to the patientfollowing stimulation and expansion.

[0225] For therapeutic purposes, CD4⁺ or CD8⁺ T cells that proliferatein response to a tumor polypeptide, polynucleotide or APC can beexpanded in number either in vitro or in vivo. Proliferation of such Tcells in vitro may be accomplished in a variety of ways. For example,the T cells can be re-exposed to a tumor polypeptide, or a short peptidecorresponding to an immunogenic portion of such a polypeptide, with orwithout the addition of T cell growth factors, such as interleukin-2,and/or stimulator cells that synthesize a tumor polypeptide.Alternatively, one or more T cells that proliferate in the presence ofthe tumor polypeptide can be expanded in number by cloning. Methods forcloning cells are well known in the art, and include limiting dilution.

[0226] T Cell Receptor Compositions

[0227] The T cell receptor (TCR) consists of 2 different, highlyvariable polypeptide chains, termed the T-cell receptor α and β chains,that are linked by a disulfide bond (Janeway, Travers, Walport.Immunobiology. Fourth Ed., 148-159, Elsevier Science Ltd/GarlandPublishing. 1999). The α/β heterodimer complexes with the invariant CD3chains at the cell membrane. This complex recognizes specific antigenicpeptides bound to MHC molecules. The enormous diversity of TCRspecificities is generated much like immunoglobulin diversity, throughsomatic gene rearrangement. The β chain genes contain over 50 variable(V), 2 diversity (D), over 10 joining (J) segments, and 2 constantregion segments (C). The α chain genes contain over 70 V segments, andover 60 J segments but no D segments, as well as one C segment. During Tcell development in the thymus, the D to J gene rearrangement of the βchain occurs, followed by the V gene segment rearrangement to the DJ.This functional VDJβ exon is transcribed and spliced to join to a Cβ.For the α chain, a Vα gene segment rearranges to a Jα gene segment tocreate the functional exon that is then transcribed and spliced to theCα. Diversity is further increased during the recombination process bythe random addition of P and N-nucleotides between the V, D, and Jsegments of the β chain and between the V and J segments in the □ chain(Janeway, Travers, Walport. Immunobiology. Fourth Ed., 98 and 150,Elsevier Science Ltd/Garland Publishing. 1999).

[0228] The present invention, in another aspect, provides TCRs specificfor a polypeptide disclosed herein, or for a variant or derivativethereof. In accordance with the present invention, polynucleotide andamino acid sequences are provided for the V-J or V-D-J junctionalregions or parts thereof for the alpha and beta chains of the T-cellreceptor which recognize tumor polypeptides described herein. Ingeneral, this aspect of the invention relates to T-cell receptors whichrecognize or bind tumor polypeptides presented in the context of MHC. Ina preferred embodiment the tumor antigens recognized by the T-cellreceptors comprise a polypeptide of the present invention. For example,cDNA encoding a TCR specific for a breast tumor peptide can be isolatedfrom T cells specific for a tumor polypeptide using standard molecularbiological and recombinant DNA techniques.

[0229] This invention further includes the T-cell receptors or analogsthereof having substantially the same function or activity as the T-cellreceptors of this invention which recognize or bind tumor polypeptides.Such receptors include, but are not limited to, a fragment of thereceptor, or a substitution, addition or deletion mutant of a T-cellreceptor provided herein. This invention also encompasses polypeptidesor peptides that are substantially homologous to the T-cell receptorsprovided herein or that retain substantially the same activity. The term“analog” includes any protein or polypeptide having an amino acidresidue sequence substantially identical to the T-cell receptorsprovided herein in which one or more residues, preferably no more than 5residues, more preferably no more than 25 residues have beenconservatively substituted with a functionally similar residue and whichdisplays the functional aspects of the T-cell receptor as describedherein.

[0230] The present invention further provides for suitable mammalianhost cells, for example, non-specific T cells, that are transfected witha polynucleotide encoding TCRs specific for a polypeptide describedherein, thereby rendering the host cell specific for the polypeptide.The α and β chains of the TCR may be contained on separate expressionvectors or alternatively, on a single expression vector that alsocontains an internal ribosome entry site (IRES) for cap-independenttranslation of the gene downstream of the IRES. Said host cellsexpressing TCRs specific for the polypeptide may be used, for example,for adoptive immunotherapy of breast cancer as discussed further below.

[0231] In further aspects of the present invention, cloned TCRs specificfor a polypeptide recited herein may be used in a kit for the diagnosisof breast cancer. For example, the nucleic acid sequence or portionsthereof, of tumor-specific TCRs can be used as probes or primers for thedetection of expression of the rearranged genes encoding the specificTCR in a biological sample. Therefore, the present invention furtherprovides for an assay for detecting messenger RNA or DNA encoding theTCR specific for a polypeptide.

[0232] Pharmaceutical Compositions

[0233] In additional embodiments, the present invention concernsformulation of one or more of the polynucleotide, polypeptide, T-cell,TCR, and/or antibody compositions disclosed herein inpharmaceutically-acceptable carriers for administration to a cell or ananimal, either alone, or in combination with one or more othermodalities of therapy.

[0234] It will be understood that, if desired, a composition asdisclosed herein may be administered in combination with other agents aswell, such as, e.g., other proteins or polypeptides or variouspharmaceutically-active agents. In fact, there is virtually no limit toother components that may also be included, given that the additionalagents do not cause a significant adverse effect upon contact with thetarget cells or host tissues. The compositions may thus be deliveredalong with various other agents as required in the particular instance.Such compositions may be purified from host cells or other biologicalsources, or alternatively may be chemically synthesized as describedherein. Likewise, such compositions may further comprise substituted orderivatized RNA or DNA compositions.

[0235] Therefore, in another aspect of the present invention,pharmaceutical compositions are provided comprising one or more of thepolynucleotide, polypeptide, antibody, TCR, and/or T-cell compositionsdescribed herein in combination with a physiologically acceptablecarrier. In certain preferred embodiments, the pharmaceuticalcompositions of the invention comprise immunogenic polynucleotide and/orpolypeptide compositions of the invention for use in prophylactic andtheraputic vaccine applications. Vaccine preparation is generallydescribed in, for example, M. F. Powell and M. J. Newman, eds., “VaccineDesign (the subunit and adjuvant approach),” Plenum Press (NY, 1995).Generally, such compositions will comprise one or more polynucleotideand/or polypeptide compositions of the present invention in combinationwith one or more immunostimulants.

[0236] It will be apparent that any of the pharmaceutical compositionsdescribed herein can contain pharmaceutically acceptable salts of thepolynucleotides and polypeptides of the invention. Such salts can beprepared, for example, from pharmaceutically acceptable non-toxic bases,including organic bases (e.g., salts of primary, secondary and tertiaryamines and basic amino acids) and inorganic bases (e.g., sodium,potassium, lithium, ammonium, calcium and magnesium salts).

[0237] In another embodiment, illustrative immunogenic compositions,e.g., vaccine compositions, of the present invention comprise DNAencoding one or more of the polypeptides as described above, such thatthe polypeptide is generated in situ. As noted above, the polynucleotidemay be administered within any of a variety of delivery systems known tothose of ordinary skill in the art. Indeed, numerous gene deliverytechniques are well known in the art, such as those described byRolland, Crit. Rev. Therap. Drug Carrier Systems 15:143-198, 1998, andreferences cited therein. Appropriate polynucleotide expression systemswill, of course, contain the necessary regulatory DNA regulatorysequences for expression in a patient (such as a suitable promoter andterminating signal). Alternatively, bacterial delivery systems mayinvolve the administration of a bacterium (such asBacillus-Calmette-Guerrin) that expresses an immunogenic portion of thepolypeptide on its cell surface or secretes such an epitope.

[0238] Therefore, in certain embodiments, polynucleotides encodingimmunogenic polypeptides described herein are introduced into suitablemammalian host cells for expression using any of a number of knownviral-based systems. In one illustrative embodiment, retrovirusesprovide a convenient and effective platform for gene delivery systems. Aselected nucleotide sequence encoding a polypeptide of the presentinvention can be inserted into a vector and packaged in retroviralparticles using techniques known in the art. The recombinant virus canthen be isolated and delivered to a subject. A number of illustrativeretroviral systems have been described (e.g., U.S. Pat. No. 5,219,740;Miller and Rosman (1989) BioTechniques 7:980-990; Miller, A. D. (1990)Human Gene Therapy 1:5-14; Scarpa et al. (1991) Virology 180:849-852;Burns et al. (1993) Proc. Natl. Acad. Sci. USA 90:8033-8037; andBoris-Lawrie and Temin (1993) Cur. Opin. Genet. Develop. 3:102-109.

[0239] In addition, a number of illustrative adenovirus-based systemshave also been described. Unlike retroviruses which integrate into thehost genome, adenoviruses persist extrachromosomally thus minimizing therisks associated with insertional mutagenesis (Haj-Ahmad and Graham(1986) J. Virol. 57:267-274; Bett et al. (1993) J. Virol. 67:5911-5921;Mittereder et al. (1994) Human Gene Therapy 5:717-729; Seth et al.(1994) J. Virol. 68:933-940; Barr et al. (1994) Gene Therapy 1:51-58;Berkner, K. L. (1988) BioTechniques 6:616-629; and Rich et al. (1993)Human Gene Therapy 4:461-476).

[0240] Various adeno-associated virus (AAV) vector systems have alsobeen developed for polynucleotide delivery. AAV vectors can be readilyconstructed using techniques well known in the art. See, e.g., U.S. Pat.Nos. 5,173,414 and 5,139,941; International Publication Nos. WO 92/01070and WO 93/03769; Lebkowski et al. (1988) Molec. Cell. Biol. 8:3988-3996;Vincent et al. (1990) Vaccines 90 (Cold Spring Harbor Laboratory Press);Carter, B. J. (1992) Current Opinion in Biotechnology 3:533-539;Muzyczka, N. (1992) Current Topics in Microbiol. and Immunol.158:97-129; Kotin, R. M. (1994) Human Gene Therapy 5:793-801; Shellingand Smith (1994) Gene Therapy 1:165-169; and Zhou et al. (1994) J. Exp.Med. 179:1867-1875.

[0241] Additional viral vectors useful for delivering thepolynucleotides encoding polypeptides of the present invention by genetransfer include those derived from the pox family of viruses, such asvaccinia virus and avian poxvirus. By way of example, vaccinia virusrecombinants expressing the novel molecules can be constructed asfollows. The DNA encoding a polypeptide is first inserted into anappropriate vector so that it is adjacent to a vaccinia promoter andflanking vaccinia DNA sequences, such as the sequence encoding thymidinekinase (TK). This vector is then used to transfect cells which aresimultaneously infected with vaccinia. Homologous recombination servesto insert the vaccinia promoter plus the gene encoding the polypeptideof interest into the viral genome. The resulting TK.sup.(−) recombinantcan be selected by culturing the cells in the presence of5-bromodeoxyuridine and picking viral plaques resistant thereto.

[0242] A vaccinia-based infection/transfection system can beconveniently used to provide for inducible, transient expression orcoexpression of one or more polypeptides described herein in host cellsof an organism. In this particular system, cells are first infected invitro with a vaccinia virus recombinant that encodes the bacteriophageT7 RNA polymerase. This polymerase displays exquisite specificity inthat it only transcribes templates bearing T7 promoters. Followinginfection, cells are transfected with the polynucleotide orpolynucleotides of interest, driven by a T7 promoter. The polymeraseexpressed in the cytoplasm from the vaccinia virus recombinanttranscribes the transfected DNA into RNA which is then translated intopolypeptide by the host translational machinery. The method provides forhigh level, transient, cytoplasmic production of large quantities of RNAand its translation products. See, e.g., Elroy-Stein and Moss, Proc.Natl. Acad. Sci. USA (1990) 87:6743-6747; Fuerst et al. Proc. Natl.Acad. Sci. USA (1986) 83:8122-8126.

[0243] Alternatively, avipoxviruses, such as the fowlpox and canarypoxviruses, can also be used to deliver the coding sequences of interest.Recombinant avipox viruses, expressing immunogens from mammalianpathogens, are known to confer protective immunity when administered tonon-avian species. The use of an Avipox vector is particularly desirablein human and other mammalian species since members of the Avipox genuscan only productively replicate in susceptible avian species andtherefore are not infective in mammalian cells. Methods for producingrecombinant Avipoxviruses are known in the art and employ geneticrecombination, as described above with respect to the production ofvaccinia viruses. See, e.g., WO 91/12882; WO 89/03429; and WO 92/03545.

[0244] Any of a number of alphavirus vectors can also be used fordelivery of polynucleotide compositions of the present invention, suchas those vectors described in U.S. Pat. Nos. 5,843,723; 6,015,686;6,008,035 and 6,015,694. Certain vectors based on Venezuelan EquineEncephalitis (VEE) can also be used, illustrative examples of which canbe found in U.S. Pat. Nos. 5,505,947 and 5,643,576.

[0245] Moreover, molecular conjugate vectors, such as the adenoviruschimeric vectors described in Michael et al. J. Biol. Chem. (1993)268:6866-6869 and Wagner et al. Proc. Natl. Acad. Sci. USA (1992)89:6099-6103, can also be used for gene delivery under the invention.

[0246] Additional illustrative information on these and other knownviral-based delivery systems can be found, for example, in Fisher-Hochet al., Proc. Natl. Acad. Sci. USA 86:317-321, 1989; Flexner et al.,Ann. N.Y. Acad. Sci. 569:86-103,1989; Flexner et al., Vaccine 8:17-21,1990; U.S. Pat. Nos. 4,603,112, 4,769,330, and 5,017,487; WO 89/01973;U.S. Pat. No. 4,777,127; GB 2,200,651; EP 0,345,242; WO 91/02805;Berkner, Biotechniques 6:616-627, 1988; Rosenfeld et al., Science252:431-434, 1991; Kolls et al., Proc. Natl. Acad. Sci. USA 91:215-219,1994; Kass-Eisler et al., Proc. Natl. Acad. Sci. USA90:11498-11502,1993; Guzman et al., Circulation 88:2838-2848, 1993; andGuzman et al., Cir. Res. 73:1202-1207, 1993.

[0247] In certain embodiments, a polynucleotide may be integrated intothe genome of a target cell. This integration may be in the specificlocation and orientation via homologous recombination (gene replacement)or it may be integrated in a random, non-specific location (geneaugmentation). In yet further embodiments, the polynucleotide may bestably maintained in the cell as a separate, episomal segment of DNA.Such polynucleotide segments or “episomes” encode sequences sufficientto permit maintenance and replication independent of or insynchronization with the host cell cycle. The manner in which theexpression construct is delivered to a cell and where in the cell thepolynucleotide remains is dependent on the type of expression constructemployed.

[0248] In another embodiment of the invention, a polynucleotide isadministered/delivered as “naked” DNA, for example as described in Ulmeret al., Science 259:1745-1749, 1993 and reviewed by Cohen, Science259:1691-1692, 1993. The uptake of naked DNA may be increased by coatingthe DNA onto biodegradable beads, which are efficiently transported intothe cells.

[0249] In still another embodiment, a composition of the presentinvention can be delivered via a particle bombardment approach, many ofwhich have been described. In one illustrative example, gas-drivenparticle acceleration can be achieved with devices such as thosemanufactured by Powderject Pharmaceuticals PLC (Oxford, UK) andPowderject Vaccines Inc. (Madison, Wis.), some examples of which aredescribed in U.S. Pat. Nos. 5,846,796; 6,010,478; 5,865,796; 5,584,807;and EP Patent No.0500 799. This approach offers a needle-free deliveryapproach wherein a dry powder formulation of microscopic particles, suchas polynucleotide or polypeptide particles, are accelerated to highspeed within a helium gas jet generated by a hand held device,propelling the particles into a target tissue of interest.

[0250] In a related embodiment, other devices and methods that may beuseful for gas-driven needle-less injection of compositions of thepresent invention include those provided by Bioject, Inc. (Portland,Oreg.), some examples of which are described in U.S. Pat. Nos.4,790,824; 5,064,413; 5,312,335; 5,383,851; 5,399,163; 5,520,639 and5,993,412.

[0251] According to another embodiment, the pharmaceutical compositionsdescribed herein will comprise one or more immunostimulants in additionto the immunogenic polynucleotide, polypeptide, antibody, T-cell, TCR,and/or APC compositions of this invention. An immunostimulant refers toessentially any substance that enhances or potentiates an immuneresponse (antibody and/or cell-mediated) to an exogenous antigen. Onepreferred type of immunostimulant comprises an adjuvant. Many adjuvantscontain a substance designed to protect the antigen from rapidcatabolism, such as aluminum hydroxide or mineral oil, and a stimulatorof immune responses, such as lipid A, Bortadella pertussis orMycobacterium tuberculosis derived proteins. Certain adjuvants arecommercially available as, for example, Freund's Incomplete Adjuvant andComplete Adjuvant (Difco Laboratories, Detroit, Mich.); Merck Adjuvant65 (Merck and Company, Inc., Rahway, N.J.); AS-2 (SmithKline Beecham,Philadelphia, Pa.); aluminum salts such as aluminum hydroxide gel (alum)or aluminum phosphate; salts of calcium, iron or zinc; an insolublesuspension of acylated tyrosine; acylated sugars; cationically oranionically derivatized polysaccharides; polyphosphazenes; biodegradablemicrospheres; monophosphoryl lipid A and quil A. Cytokines, such asGM-CSF, interleukin-2,-7,-12, and other like growth factors, may also beused as adjuvants.

[0252] Within certain embodiments of the invention, the adjuvantcomposition is preferably one that induces an immune responsepredominantly of the Th1 type. High levels of Th1-type cytokines (e.g.,IFN-γ, TNFα, IL-2 and IL-12) tend to favor the induction of cellmediated immune responses to an administered antigen. In contrast, highlevels of Th2-type cytokines (e.g., IL-4, IL-5, IL-6 and IL-10) tend tofavor the induction of humoral immune responses. Following applicationof a vaccine as provided herein, a patient will support an immuneresponse that includes Th1- and Th2-type responses. Within a preferredembodiment, in which a response is predominantly Th1-type, the level ofTh1-type cytokines will increase to a greater extent than the level ofTh2-type cytokines. The levels of these cytokines may be readilyassessed using standard assays. For a review of the families ofcytokines, see Mosmann and Coffman, Ann. Rev. Immunol. 7:145-173, 1989.

[0253] Certain preferred adjuvants for eliciting a predominantlyTh1-type response include, for example, a combination of monophosphoryllipid A, preferably 3-de-O-acylated monophosphoryl lipid A, togetherwith an aluminum salt. MPL® adjuvants are available from CorixaCorporation (Seattle, Wash.; see, for example, U.S. Pat. Nos. 4,436,727;4,877,611; 4,866,034 and 4,912,094). CpG-containing oligonucleotides (inwhich the CpG dinucleotide is unmethylated) also induce a predominantlyTh1 response. Such oligonucleotides are well known and are described,for example, in WO 96/02555, WO 99/33488 and U.S. Pat. Nos. 6,008,200and 5,856,462. Immunostimulatory DNA sequences are also described, forexample, by Sato et al., Science 273:352, 1996. Another preferredadjuvant comprises a saponin, such as Quil A, or derivatives thereof,including QS21 and QS7 (Aquila Biopharmaceuticals Inc., Framingham,Mass.); Escin; Digitonin; or Gypsophila or Chenopodium quinoa saponins.Other preferred formulations include more than one saponin in theadjuvant combinations of the present invention, for example combinationsof at least two of the following group comprising QS21, QS7, Quil A,β-escin, or digitonin.

[0254] Alternatively the saponin formulations may be combined withvaccine vehicles composed of chitosan or other polycationic polymers,polylactide and polylactide-co-glycolide particles, poly-N-acetylglucosamine-based polymer matrix, particles composed of polysaccharidesor chemically modified polysaccharides, liposomes and lipid-basedparticles, particles composed of glycerol monoesters, etc. The saponinsmay also be formulated in the presence of cholesterol to formparticulate structures such as liposomes or ISCOMs. Furthermore, thesaponins may be formulated together with a polyoxyethylene ether orester, in either a non-particulate solution or suspension, or in aparticulate structure such as a paucilamelar liposome or ISCOM. Thesaponins may also be formulated with excipients such as Carbopol^(R) toincrease viscosity, or may be formulated in a dry powder form with apowder excipient such as lactose.

[0255] In one preferred embodiment, the adjuvant system includes thecombination of a monophosphoryl lipid A and a saponin derivative, suchas the combination of QS21 and 3D-MPL® adjuvant, as described in WO94/00153, or a less reactogenic composition where the QS21 is quenchedwith cholesterol, as described in WO 96/33739. Other preferredformulations comprise an oil-in-water emulsion and tocopherol. Anotherparticularly preferred adjuvant formulation employing QS21, 3D-MPL®adjuvant and tocopherol in an oil-in-water emulsion is described in WO95/17210.

[0256] Another enhanced adjuvant system involves the combination of aCpG-containing oligonucleotide and a saponin derivative particularly thecombination of CpG and QS21 is disclosed in WO 00/09159. Preferably theformulation additionally comprises an oil in water emulsion andtocopherol.

[0257] Additional illustrative adjuvants for use in the pharmaceuticalcompositions of the invention include Montanide ISA 720 (Seppic,France), SAF (Chiron, Calif., United States), ISCOMS (CSL), MF-59(Chiron), the SBAS series of adjuvants (e.g., SBAS-2 or SBAS-4,available from SmithKline Beecham, Rixensart, Belgium), Detox(Enhanzyn®) (Corixa, Hamilton, Mont.), RC-529 (Corixa, Hamilton, Mont.)and other aminoalkyl glucosaminide 4-phosphates (AGPs), such as thosedescribed in pending U.S. patent application Ser. Nos. 08/853,826 and09/074,720, the disclosures of which are incorporated herein byreference in their entireties, and polyoxyethylene ether adjuvants suchas those described in WO 99/52549A1.

[0258] Other preferred adjuvants include adjuvant molecules of thegeneral formula

HO(CH₂CH₂O)_(n)-A-R,   (I)

[0259] wherein, n is 1-50, A is a bond or —C(O)—, R is C₁₋₅₀ alkyl orPhenyl C₁₋₅₀ alkyl.

[0260] One embodiment of the present invention consists of a vaccineformulation comprising a polyoxyethylene ether of general formula (I),wherein n is between 1 and 50, preferably 4-24, most preferably 9; the Rcomponent is C₁₋₅₀, preferably C₄-C₂₀ alkyl and most preferably C₁₂alkyl, and A is a bond. The concentration of the polyoxyethylene ethersshould be in the range 0.1-20%, preferably from 0.1-10%, and mostpreferably in the range 0.1-1%. Preferred polyoxyethylene ethers areselected from the following group: polyoxyethylene-9-lauryl ether,polyoxyethylene-9-steoryl ether, polyoxyethylene-8-steoryl ether,polyoxyethylene-4-lauryl ether, polyoxyethylene-35-lauryl ether, andpolyoxyethylene-23-lauryl ether. Polyoxyethylene ethers such aspolyoxyethylene lauryl ether are described in the Merck index (12^(th)edition: entry 7717). These adjuvant molecules are described in WO99/52549.

[0261] The polyoxyethylene ether according to the general formula (I)above may, if desired, be combined with another adjuvant. For example, apreferred adjuvant combination is preferably with CpG as described inthe pending UK patent application GB 9820956.2.

[0262] According to another embodiment of this invention, an immunogeniccomposition described herein is delivered to a host via antigenpresenting cells (APCs), such as dendritic cells, macrophages, B cells,monocytes and other cells that may be engineered to be efficient APCs.Such cells may, but need not, be genetically modified to increase thecapacity for presenting the antigen, to improve activation and/ormaintenance of the T cell response, to have anti-tumor effects per seand/or to be immunologically compatible with the receiver (i.e., matchedHLA haplotype). APCs may generally be isolated from any of a variety ofbiological fluids and organs, including tumor and peritumoral tissues,and may be autologous, allogeneic, syngeneic or xenogeneic cells.

[0263] Certain preferred embodiments of the present invention usedendritic cells or progenitors thereof as antigen-presenting cells.Dendritic cells are highly potent APCs (Banchereau and Steinman, Nature392:245-251, 1998) and have been shown to be effective as aphysiological adjuvant for eliciting prophylactic or therapeuticantitumor immunity (see Timmerman and Levy, Ann. Rev. Med.50:507-529,1999). In general, dendritic cells may be identified based ontheir typical shape (stellate in situ, with marked cytoplasmic processes(dendrites) visible in vitro), their ability to take up, process andpresent antigens with high efficiency and their ability to activatenaïve T cell responses. Dendritic cells may, of course, be engineered toexpress specific cell-surface receptors or ligands that are not commonlyfound on dendritic cells in vivo or ex vivo, and such modified dendriticcells are contemplated by the present invention. As an alternative todendritic cells, secreted vesicles antigen-loaded dendritic cells(called exosomes) may be used within a vaccine (see Zitvogel et al.,Nature Med. 4:594-600, 1998).

[0264] Dendritic cells and progenitors may be obtained from peripheralblood, bone marrow, tumor-infiltrating cells, peritumoraltissues-infiltrating cells, lymph nodes, spleen, skin, umbilical cordblood or any other suitable tissue or fluid. For example, dendriticcells may be differentiated ex vivo by adding a combination of cytokinessuch as GM-CSF, IL-4, IL-13 and/or TNFα to cultures of monocytesharvested from peripheral blood. Alternatively, CD34 positive cellsharvested from peripheral blood, umbilical cord blood or bone marrow maybe differentiated into dendritic cells by adding to the culture mediumcombinations of GM-CSF, IL-3, TNFα, CD40 ligand, LPS, fit3 ligand and/orother compound(s) that induce differentiation, maturation andproliferation of dendritic cells.

[0265] Dendritic cells are conveniently categorized as “immature” and“mature” cells, which allows a simple way to discriminate between twowell characterized phenotypes. However, this nomenclature should not beconstrued to exclude all possible intermediate stages ofdifferentiation. Immature dendritic cells are characterized as APC witha high capacity for antigen uptake and processing, which correlates withthe high expression of Fcγ receptor and mannose receptor. The maturephenotype is typically characterized by a lower expression of thesemarkers, but a high expression of cell surface molecules responsible forT cell activation such as class I and class II MHC, adhesion molecules(e.g., CD54 and CD11) and costimulatory molecules (e.g., CD40, CD80,CD86 and 4-1BB).

[0266] APCs may generally be transfected with a polynucleotide of theinvention (or portion or other variant thereof) such that the encodedpolypeptide, or an immunogenic portion thereof, is expressed on the cellsurface. Such transfection may take place ex vivo, and a pharmaceuticalcomposition comprising such transfected cells may then be used fortherapeutic purposes, as described herein. Alternatively, a genedelivery vehicle that targets a dendritic or other antigen presentingcell may be administered to a patient, resulting in transfection thatoccurs in vivo. In vivo and ex vivo transfection of dendritic cells, forexample, may generally be performed using any methods known in the art,such as those described in WO 97/24447, or the gene gun approachdescribed by Mahvi et al., Immunology and cell Biology 75:456-460,1997.Antigen loading of dendritic cells may be achieved by incubatingdendritic cells or progenitor cells with the tumor polypeptide, DNA(naked or within a plasmid vector) or RNA; or with antigen-expressingrecombinant bacterium or viruses (e.g., vaccinia, fowlpox, adenovirus orlentivirus vectors). Prior to loading, the polypeptide may be covalentlyconjugated to an immunological partner that provides T cell help (e.g.,a carrier molecule). Alternatively, a dendritic cell may be pulsed witha non-conjugated immunological partner, separately or in the presence ofthe polypeptide.

[0267] While any suitable carrier known to those of ordinary skill inthe art may be employed in the pharmaceutical compositions of thisinvention, the type of carrier will typically vary depending on the modeof administration. Compositions of the present invention may beformulated for any appropriate manner of administration, including forexample, topical, oral, nasal, mucosal, intravenous, intracranial,intraperitoneal, subcutaneous and intramuscular administration.

[0268] Carriers for use within such pharmaceutical compositions arebiocompatible, and may also be biodegradable. In certain embodiments,the formulation preferably provides a relatively constant level ofactive component release. In other embodiments, however, a more rapidrate of release immediately upon administration may be desired. Theformulation of such compositions is well within the level of ordinaryskill in the art using known techniques. Illustrative carriers useful inthis regard include microparticles of poly(lactide-co-glycolide),polyacrylate, latex, starch, cellulose, dextran and the like. Otherillustrative delayed-release carriers include supramolecular biovectors,which comprise a non-liquid hydrophilic core (e.g., a cross-linkedpolysaccharide or oligosaccharide) and, optionally, an external layercomprising an amphiphilic compound, such as a phospholipid (see e.g.,U.S. Pat. No. 5,151,254 and PCT applications WO 94/20078, WO/94/23701and WO 96/06638). The amount of active compound contained within asustained release formulation depends upon the site of implantation, therate and expected duration of release and the nature of the condition tobe treated or prevented.

[0269] In another illustrative embodiment, biodegradable microspheres(e.g., polylactate polyglycolate) are employed as carriers for thecompositions of this invention. Suitable biodegradable microspheres aredisclosed, for example, in U.S. Pat. Nos. 4,897,268; 5,075,109;5,928,647; 5,811,128; 5,820,883; 5,853,763; 5,814,344, 5,407,609 and5,942,252. Modified hepatitis B core protein carrier systems, such asdescribed in WO/99 40934, and references cited therein, will also beuseful for many applications. Another illustrative carrier/deliverysystem employs a carrier comprising particulate-protein complexes, suchas those described in U.S. Pat. No. 5,928,647, which are capable ofinducing a class I-restricted cytotoxic T lymphocyte responses in ahost.

[0270] In another illustrative embodiment, calcium phosphate coreparticles are employed as carriers, vaccine adjuvants, or as controlledrelease matrices for the compositions of this invention. Exemplarycalcium phosphate particles are disclosed, for example, in publishedpatent application No. WO/0046147.

[0271] The pharmaceutical compositions of the invention will oftenfurther comprise one or more buffers (e.g., neutral buffered saline orphosphate buffered saline), carbohydrates (e.g., glucose, mannose,sucrose or dextrans), mannitol, proteins, polypeptides or amino acidssuch as glycine, antioxidants, bacteriostats, chelating agents such asEDTA or glutathione, adjuvants (e.g., aluminum hydroxide), solutes thatrender the formulation isotonic, hypotonic or weakly hypertonic with theblood of a recipient, suspending agents, thickening agents and/orpreservatives. Alternatively, compositions of the present invention maybe formulated as a lyophilizate.

[0272] The pharmaceutical compositions described herein may be presentedin unit-dose or multi-dose containers, such as sealed ampoules or vials.Such containers are typically sealed in such a way to preserve thesterility and stability of the formulation until use. In general,formulations may be stored as suspensions, solutions or emulsions inoily or aqueous vehicles. Alternatively, a pharmaceutical compositionmay be stored in a freeze-dried condition requiring only the addition ofa sterile liquid carrier immediately prior to use.

[0273] The development of suitable dosing and treatment regimens forusing the particular compositions described herein in a variety oftreatment regimens, including e.g., oral, parenteral, intravenous,intranasal, and intramuscular administration and formulation, is wellknown in the art, some of which are briefly discussed below for generalpurposes of illustration.

[0274] In certain applications, the pharmaceutical compositionsdisclosed herein may be delivered via oral administration to an animal.As such, these compositions may be formulated with an inert diluent orwith an assimilable edible carrier, or they may be enclosed in hard- orsoft-shell gelatin capsule, or they may be compressed into tablets, orthey may be incorporated directly with the food of the diet.

[0275] The active compounds may even be incorporated with excipients andused in the form of ingestible tablets, buccal tables, troches,capsules, elixirs, suspensions, syrups, wafers, and the like (see, forexample, Mathiowitz et al., Nature Mar. 27, 1997;386(6623):410-4; Hwanget al., Crit Rev Ther Drug Carrier Syst 1998;15(3):243-84; U.S. Pat. No.5,641,515; U.S. Pat. No. 5,580,579 and U.S. Pat. No. 5,792,451)U.S.Tablets, troches, pills, capsules and the like may also contain any of avariety of additional components, for example, a binder, such as gumtragacanth, acacia, cornstarch, or gelatin; excipients, such asdicalcium phosphate; a disintegrating agent, such as corn starch, potatostarch, alginic acid and the like; a lubricant, such as magnesiumstearate; and a sweetening agent, such as sucrose, lactose or saccharinmay be added or a flavoring agent, such as peppermint, oil ofwintergreen, or cherry flavoring. When the dosage unit form is acapsule, it may contain, in addition to materials of the above type, aliquid carrier. Various other materials may be present as coatings or tootherwise modify the physical form of the dosage unit. For instance,tablets, pills, or capsules may be coated with shellac, sugar, or both.Of course, any material used in preparing any dosage unit form should bepharmaceutically pure and substantially non-toxic in the amountsemployed. In addition, the active compounds may be incorporated intosustained-release preparation and formulations.

[0276] Typically, these formulations will contain at least about 0.1% ofthe active compound or more, although the percentage of the activeingredient(s) may, of course, be varied and may conveniently be betweenabout 1 or 2% and about 60% or 70% or more of the weight or volume ofthe total formulation. Naturally, the amount of active compound(s) ineach therapeutically useful composition may be prepared is such a waythat a suitable dosage will be obtained in any given unit dose of thecompound. Factors such as solubility, bioavailability, biologicalhalf-life, route of administration, product shelf life, as well as otherpharmacological considerations will be contemplated by one skilled inthe art of preparing such pharmaceutical formulations, and as such, avariety of dosages and treatment regimens may be desirable.

[0277] For oral administration the compositions of the present inventionmay alternatively be incorporated with one or more excipients in theform of a mouthwash, dentifrice, buccal tablet, oral spray, orsublingual orally-administered formulation. Alternatively, the activeingredient may be incorporated into an oral solution such as onecontaining sodium borate, glycerin and potassium bicarbonate, ordispersed in a dentifrice, or added in a therapeutically-effectiveamount to a composition that may include water, binders, abrasives,flavoring agents, foaming agents, and humectants. Alternatively thecompositions may be fashioned into a tablet or solution form that may beplaced under the tongue or otherwise dissolved in the mouth.

[0278] In certain circumstances it will be desirable to deliver thepharmaceutical compositions disclosed herein parenterally,intravenously, intramuscularly, or even intraperitoneally. Suchapproaches are well known to the skilled artisan, some of which arefurther described, for example, in U.S. Pat. No. 5,543,158; U.S. Pat.No. 5,641,515 and U.S. Pat. No. 5,399,363. In certain embodiments,solutions of the active compounds as free base or pharmacologicallyacceptable salts may be prepared in water suitably mixed with asurfactant, such as hydroxypropylcellulose. Dispersions may also beprepared in glycerol, liquid polyethylene glycols, and mixtures thereofand in oils. Under ordinary conditions of storage and use, thesepreparations generally will contain a preservative to prevent the growthof microorganisms.

[0279] Illustrative pharmaceutical forms suitable for injectable useinclude sterile aqueous solutions or dispersions and sterile powders forthe extemporaneous preparation of sterile injectable solutions ordispersions (for example, see U.S. Pat. No. 5,466,468). In all cases theform must be sterile and must be fluid to the extent that easysyringability exists. It must be stable under the conditions ofmanufacture and storage and must be preserved against the contaminatingaction of microorganisms, such as bacteria and fungi. The carrier can bea solvent or dispersion medium containing, for example, water, ethanol,polyol (e.g., glycerol, propylene glycol, and liquid polyethyleneglycol, and the like), suitable mixtures thereof, and/or vegetable oils.Proper fluidity may be maintained, for example, by the use of a coating,such as lecithin, by the maintenance of the required particle size inthe case of dispersion and/or by the use of surfactants. The preventionof the action of microorganisms can be facilitated by variousantibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In manycases, it will be preferable to include isotonic agents, for example,sugars or sodium chloride. Prolonged absorption of the injectablecompositions can be brought about by the use in the compositions ofagents delaying absorption, for example, aluminum monostearate andgelatin.

[0280] In one embodiment, for parenteral administration in an aqueoussolution, the solution should be suitably buffered if necessary and theliquid diluent first rendered isotonic with sufficient saline orglucose. These particular aqueous solutions are especially suitable forintravenous, intramuscular, subcutaneous and intraperitonealadministration. In this connection, a sterile aqueous medium that can beemployed will be known to those of skill in the art in light of thepresent disclosure. For example, one dosage may be dissolved in 1 ml ofisotonic NaCl solution and either added to 1000 ml of hypodermoclysisfluid or injected at the proposed site of infusion, (see for example,“Remington's Pharmaceutical Sciences” 15th Edition, pages 1035-1038 and1570-1580). Some variation in dosage will necessarily occur depending onthe condition of the subject being treated. Moreover, for humanadministration, preparations will of course preferably meet sterility,pyrogenicity, and the general safety and purity standards as required byFDA Office of Biologics standards.

[0281] In another embodiment of the invention, the compositionsdisclosed herein may be formulated in a neutral or salt form.Illustrative pharmaceutically-acceptable salts include the acid additionsalts (formed with the free amino groups of the protein) and which areformed with inorganic acids such as, for example, hydrochloric orphosphoric acids, or such organic acids as acetic, oxalic, tartaric,mandelic, and the like. Salts formed with the free carboxyl groups canalso be derived from inorganic bases such as, for example, sodium,potassium, ammonium, calcium, or ferric hydroxides, and such organicbases as isopropylamine, trimethylamine, histidine, procaine and thelike. Upon formulation, solutions will be administered in a mannercompatible with the dosage formulation and in such amount as istherapeutically effective.

[0282] The carriers can further comprise any and all solvents,dispersion media, vehicles, coatings, diluents, antibacterial andantifungal agents, isotonic and absorption delaying agents, buffers,carrier solutions, suspensions, colloids, and the like. The use of suchmedia and agents for pharmaceutical active substances is well known inthe art. Except insofar as any conventional media or agent isincompatible with the active ingredient, its use in the therapeuticcompositions is contemplated. Supplementary active ingredients can alsobe incorporated into the compositions. The phrase“pharmaceutically-acceptable” refers to molecular entities andcompositions that do not produce an allergic or similar untowardreaction when administered to a human.

[0283] In certain embodiments, the pharmaceutical compositions may bedelivered by intranasal sprays, inhalation, and/or other aerosoldelivery vehicles. Methods for delivering genes, nucleic acids, andpeptide compositions directly to the lungs via nasal aerosol sprays hasbeen described, e.g., in U.S. Pat. No. 5,756,353 and U.S. Pat. No.5,804,212. Likewise, the delivery of drugs using intranasalmicroparticle resins (Takenaga et al., J Controlled Release Mar. 2,1998;52(1-2):81-7) and lysophosphatidyl-glycerol compounds (U.S. Pat.No. 5,725,871) are also well-known in the pharmaceutical arts. Likewise,illustrative transmucosal drug delivery in the form of apolytetrafluoroetheylene support matrix is described in U.S. Pat. No.5,780,045.

[0284] In certain embodiments, liposomes, nanocapsules, microparticles,lipid particles, vesicles, and the like, are used for the introductionof the compositions of the present invention into suitable hostcells/organisms. In particular, the compositions of the presentinvention may be formulated for delivery either encapsulated in a lipidparticle, a liposome, a vesicle, a nanosphere, or a nanoparticle or thelike. Alternatively, compositions of the present invention can be bound,either covalently or non-covalently, to the surface of such carriervehicles.

[0285] The formation and use of liposome and liposome-like preparationsas potential drug carriers is generally known to those of skill in theart (see for example, Lasic, Trends Biotechnol July 1998;16(7):307-21;Takakura, Nippon Rinsho March 1998;56(3):691-5; Chandran et al., IndianJ Exp Biol. August 1997;35(8):801-9; Margalit, Crit Rev Ther DrugCarrier Syst. 1995;12(2-3):233-61; U.S. Pat. No. 5,567,434; U.S. Pat.No. 5,552,157; U.S. Pat. No. 5,565,213; U.S. Pat. No. 5,738,868 and U.S.Pat. No. 5,795,587, each specifically incorporated herein by referencein its entirety).

[0286] Liposomes have been used successfully with a number of cell typesthat are normally difficult to transfect by other procedures, includingT cell suspensions, primary hepatocyte cultures and PC 12 cells(Renneisen et al., J Biol Chem. Sep. 25, 1990;265(27):16337-42; Mulleret al., DNA Cell Biol. April 1990;9(3):221-9). In addition, liposomesare free of the DNA length constraints that are typical of viral-baseddelivery systems. Liposomes have been used effectively to introducegenes, various drugs, radiotherapeutic agents, enzymes, viruses,transcription factors, allosteric effectors and the like, into a varietyof cultured cell lines and animals. Furthermore, he use of liposomesdoes not appear to be associated with autoimmune responses orunacceptable toxicity after systemic delivery.

[0287] In certain embodiments, liposomes are formed from phospholipidsthat are dispersed in an aqueous medium and spontaneously formmultilamellar concentric bilayer vesicles (also termed multilamellarvesicles (MLVs).

[0288] Alternatively, in other embodiments, the invention provides forpharmaceutically-acceptable nanocapsule formulations of the compositionsof the present invention. Nanocapsules can generally entrap compounds ina stable and reproducible way (see, for example, Quintanar-Guerrero etal., Drug Dev Ind Pharm. December 1998 24(12):1113-28). To avoid sideeffects due to intracellular polymeric overloading, such ultrafineparticles (sized around 0.1 μm) may be designed using polymers able tobe degraded in vivo. Such particles can be made as described, forexample, by Couvreur et al., Crit Rev Ther Drug Carrier Syst.1988;5(1):1-20; zur Muhlen et al., Eur J Pharm Biopharm. March1998;45(2):149-55; Zambaux et al. J Controlled Release. Jan. 2,1998;50(1-3):31-40; and U.S. Pat. No. 5,145,684.

[0289] Cancer Therapeutic Methods

[0290] Immunologic approaches to cancer therapy are based on therecognition that cancer cells can often evade the body's defensesagainst aberrant or foreign cells and molecules, and that these defensesmight be therapeutically stimulated to regain the lost ground, e.g.,pgs. 623-648 in Klein, Immunology (Wiley-lnterscience, New York, 1982).Numerous recent observations that various immune effectors can directlyor indirectly inhibit growth of tumors has led to renewed interest inthis approach to cancer therapy, e.g., Jager, et al., Oncology2001;60(1):1-7; Renner, et al., Ann Hematol December 2000;79(12):651-9.

[0291] Four-basic cell types whose function has been associated withantitumor cell immunity and the elimination of tumor cells from the bodyare: i) B-lymphocytes which secrete immunoglobulins into the bloodplasma for identifying and labeling the nonself invader cells; ii)monocytes which secrete the complement proteins that are responsible forlysing and processing the immunoglobulin-coated target invader cells;iii) natural killer lymphocytes having two mechanisms for thedestruction of tumor cells, antibody-dependent cellular cytotoxicity andnatural killing; and iv) T-lymphocytes possessing antigen-specificreceptors and having the capacity to recognize a tumor cell carryingcomplementary marker molecules (Schreiber, H., 1989, in FundamentalImmunology (ed.) W. E. Paul, pp. 923-955).

[0292] Cancer immunotherapy generally focuses on inducing humoral immuneresponses, cellular immune responses, or both. Moreover, it is wellestablished that induction of CD4⁺ T helper cells is necessary in orderto secondarily induce either antibodies or cytotoxic CD8⁺ T cells.Polypeptide antigens that are selective or ideally specific for cancercells, particularly breast cancer cells, offer a powerful approach forinducing immune responses against breast cancer, and are an importantaspect of the present invention.

[0293] Therefore, in further aspects of the present invention, thepharmaceutical compositions described herein may be used to stimulate animmune response against cancer, particularly for the immunotherapy ofbreast cancer. Within such methods, the pharmaceutical compositionsdescribed herein are administered to a patient, typically a warm-bloodedanimal, preferably a human. A patient may or may not be afflicted withcancer. Pharmaceutical compositions and vaccines may be administeredeither prior to or following surgical removal of primary tumors and/ortreatment such as administration of radiotherapy or conventionalchemotherapeutic drugs. As discussed above, administration of thepharmaceutical compositions may be by any suitable method, includingadministration by intravenous, intraperitoneal, intramuscular,subcutaneous, intranasal, intradermal, anal, vaginal, topical and oralroutes.

[0294] Within certain embodiments, immunotherapy may be activeimmunotherapy, in which treatment relies on the in vivo stimulation ofthe endogenous host immune system to react against tumors with theadministration of immune response-modifying agents (such as polypeptidesand polynucleotides as provided herein).

[0295] Within other embodiments, immunotherapy may be passiveimmunotherapy, in which treatment involves the delivery of agents withestablished tumor-immune reactivity (such as effector cells orantibodies) that can directly or indirectly mediate antitumor effectsand does not necessarily depend on an intact host immune system.Examples of effector cells include T cells as discussed above, Tlymphocytes (such as CD8⁺ cytotoxic T lymphocytes and CD4⁺ T-helpertumor-infiltrating lymphocytes), killer cells (such as Natural Killercells and lymphokine-activated killer cells), B cells andantigen-presenting cells (such as dendritic cells and macrophages)expressing a polypeptide provided herein. T cell receptors and antibodyreceptors specific for the polypeptides recited herein may be cloned,expressed and transferred into other vectors or effector cells foradoptive immunotherapy. The polypeptides provided herein may also beused to generate antibodies or anti-idiotypic antibodies (as describedabove and in U.S. Pat. No. 4,918,164) for passive immunotherapy.

[0296] Monoclonal antibodies may be labeled with any of a variety oflabels for desired selective usages in detection, diagnostic assays ortherapeutic applications (as described in U.S. Pat. Nos. 6,090,365;6,015,542; 5,843,398; 5,595,721; and 4,708,930, hereby incorporated byreference in their entirety as if each was incorporated individually).In each case, the binding of the labelled monoclonal antibody to thedeterminant site of the antigen will signal detection or delivery of aparticular therapeutic agent to the antigenic determinant on thenon-normal cell. A further object of this invention is to provide thespecific monoclonal antibody suitably labelled for achieving suchdesired selective usages thereof.

[0297] Effector cells may generally be obtained in sufficient quantitiesfor adoptive immunotherapy by growth in vitro, as described herein.Culture conditions for expanding single antigen-specific effector cellsto several billion in number with retention of antigen recognition invivo are well known in the art. Such in vitro culture conditionstypically use intermittent stimulation with antigen, often in thepresence of cytokines (such as IL-2) and non-dividing feeder cells. Asnoted above, immunoreactive polypeptides as provided herein may be usedto rapidly expand antigen-specific T cell cultures in order to generatea sufficient number of cells for immunotherapy. In particular,antigen-presenting cells, such as dendritic, macrophage, monocyte,fibroblast and/or B cells, may be pulsed with immunoreactivepolypeptides or transfected with one or more polynucleotides usingstandard techniques well known in the art. For example,antigen-presenting cells can be transfected with a polynucleotide havinga promoter appropriate for increasing expression in a recombinant virusor other expression system. Cultured effector cells for use in therapymust be able to grow and distribute widely, and to survive long term invivo. Studies have shown that cultured effector cells can be induced togrow in vivo and to survive long term in substantial numbers by repeatedstimulation with antigen supplemented with IL-2 (see, for example,Cheever et al., Immunological Reviews 157:177, 1997).

[0298] Alternatively, a vector expressing a polypeptide recited hereinmay be introduced into antigen presenting cells taken from a patient andclonally propagated ex vivo for transplant back into the same patient.Transfected cells may be reintroduced into the patient using any meansknown in the art, preferably in sterile form by intravenous,intracavitary, intraperitoneal or intratumor administration.

[0299] Routes and frequency of administration of the therapeuticcompositions described herein, as well as dosage, will vary fromindividual to individual, and may be readily established using standardtechniques. In general, the pharmaceutical compositions and vaccines maybe administered by injection (e.g., intracutaneous, intramuscular,intravenous or subcutaneous), intranasally (e.g., by aspiration) ororally. Preferably, between 1 and 10 doses may be administered over a 52week period. Preferably, 6 doses are administered, at intervals of 1month, and booster vaccinations may be given periodically thereafter.Alternate protocols may be appropriate for individual patients. Asuitable dose is an amount of a compound that, when administered asdescribed above, is capable of promoting an anti-tumor immune response,and is at least 10-50% above the basal (i.e., untreated) level. Suchresponse can be monitored by measuring the anti-tumor antibodies in apatient or by vaccine-dependent generation of cytolytic effector cellscapable of killing the patient's tumor cells in vitro. Such vaccinesshould also be capable of causing an immune response that leads to animproved clinical outcome (e.g., more frequent remissions, complete orpartial or longer disease-free survival) in vaccinated patients ascompared to non-vaccinated patients. In general, for pharmaceuticalcompositions and vaccines comprising one or more polypeptides, theamount of each polypeptide present in a dose ranges from about 25 μg to5 mg per kg of host. Suitable dose sizes will vary with the size of thepatient, but will typically range from about 0.1 mL to about 5 mL.

[0300] In general, an appropriate dosage and treatment regimen providesthe active compound(s) in an amount sufficient to provide therapeuticand/or prophylactic benefit. Such a response can be monitored byestablishing an improved clinical outcome (e.g., more frequentremissions, complete or partial, or longer disease-free survival) intreated patients as compared to non-treated patients. Increases inpreexisting immune responses to a tumor protein generally correlate withan improved clinical outcome. Such immune responses may generally beevaluated using standard proliferation, cytotoxicity or cytokine assays,which may be performed using samples obtained from a patient before andafter treatment.

[0301] Cancer Detection and Diagnostic compositions, Methods and Kits

[0302] In general, a cancer may be detected in a patient based on thepresence of one or more breast tumor proteins and/or polynucleotidesencoding such proteins in a biological sample (for example, blood, sera,sputum urine and/or tumor biopsies) obtained from the patient. In otherwords, such proteins may be used as markers to indicate the presence orabsence of a cancer such as breast cancer. In addition, such proteinsmay be useful for the detection of other cancers. The binding agentsprovided herein generally permit detection of the level of antigen thatbinds to the agent in the biological sample.

[0303] Polynucleotide primers and probes may be used to detect the levelof mRNA encoding a tumor protein, which is also indicative of thepresence or absence of a cancer. In general, a tumor sequence should bepresent at a level that is at least two-fold, preferably three-fold, andmore preferably five-fold or higher in tumor tissue than in normaltissue of the same type from which the tumor arose. Expression levels ofa particular tumor sequence in tissue types different from that in whichthe tumor arose are irrelevant in certain diagnostic embodiments sincethe presence of tumor cells can be confirmed by observation ofpredetermined differential expression levels, e.g., 2-fold, 5-fold, etc,in tumor tissue to expression levels in normal tissue of the same type.

[0304] Other differential expression patterns can be utilizedadvantageously for diagnostic purposes. For example, in one aspect ofthe invention, overexpression of a tumor sequence in tumor tissue andnormal tissue of the same type, but not in other normal tissue types,e.g., PBMCs, can be exploited diagnostically. In this case, the presenceof metastatic tumor cells, for example in a sample taken from thecirculation or some other tissue site different from that in which thetumor arose, can be identified and/or confirmed by detecting expressionof the tumor sequence in the sample, for example using RT-PCR analysis.In many instances, it will be desired to enrich for tumor cells in thesample of interest, e.g., PBMCs, using cell capture or other liketechniques.

[0305] There are a variety of assay formats known to those of ordinaryskill in the art for using a binding agent to detect polypeptide markersin a sample. See, e.g., Harlow and Lane, Antibodies: A LaboratoryManual, Cold Spring Harbor Laboratory, 1988. In general, the presence orabsence of a cancer in a patient may be determined by (a) contacting abiological sample obtained from a patient with a binding agent; (b)detecting in the sample a level of polypeptide that binds to the bindingagent; and (c) comparing the level of polypeptide with a predeterminedcut-off value.

[0306] In a preferred embodiment, the assay involves the use of bindingagent immobilized on a solid support to bind to and remove thepolypeptide from the remainder of the sample. The bound polypeptide maythen be detected using a detection reagent that contains a reportergroup and specifically binds to the binding agent/polypeptide complex.Such detection reagents may comprise, for example, a binding agent thatspecifically binds to the polypeptide or an antibody or other agent thatspecifically binds to the binding agent, such as an anti-immunoglobulin,protein G, protein A or a lectin. Alternatively, a competitive assay maybe utilized, in which a polypeptide is labeled with a reporter group andallowed to bind to the immobilized binding agent after incubation of thebinding agent with the sample. The extent to which components of thesample inhibit the binding of the labeled polypeptide to the bindingagent is indicative of the reactivity of the sample with the immobilizedbinding agent. Suitable polypeptides for use within such assays includefull length breast tumor proteins and polypeptide portions thereof towhich the binding agent binds, as described above.

[0307] The solid support may be any material known to those of ordinaryskill in the art to which the tumor protein may be attached. Forexample, the solid support may be a test well in a microtiter plate or anitrocellulose or other suitable membrane. Alternatively, the supportmay be a bead or disc, such as glass, fiberglass, latex or a plasticmaterial such as polystyrene or polyvinylchloride. The support may alsobe a magnetic particle or a fiber optic sensor, such as those disclosed,for example, in U.S. Pat. No. 5,359,681. The binding agent may beimmobilized on the solid support using a variety of techniques known tothose of skill in the art, which are amply described in the patent andscientific literature. In the context of the present invention, the term“immobilization” refers to both noncovalent association, such asadsorption, and covalent attachment (which may be a direct linkagebetween the agent and functional groups on the support or may be alinkage by way of a cross-linking agent). Immobilization by adsorptionto a well in a microtiter plate or to a membrane is preferred. In suchcases, adsorption may be achieved by contacting the binding agent, in asuitable buffer, with the solid support for a suitable amount of time.The contact time varies with temperature, but is typically between about1 hour and about 1 day. In general, contacting a well of a plasticmicrotiter plate (such as polystyrene or polyvinylchloride) with anamount of binding agent ranging from about 10 ng to about 10 μg, andpreferably about 100 ng to about 1 μg, is sufficient to immobilize anadequate amount of binding agent.

[0308] Covalent attachment of binding agent to a solid support maygenerally be achieved by first reacting the support with a bifunctionalreagent that will react with both the support and a functional group,such as a hydroxyl or amino group, on the binding agent. For example,the binding agent may be covalently attached to supports having anappropriate polymer coating using benzoquinone or by condensation of analdehyde group on the support with an amine and an active hydrogen onthe binding partner (see, e.g., Pierce Immunotechnology Catalog andHandbook, 1991, at A12-A13).

[0309] In certain embodiments, the assay is a two-antibody sandwichassay. This assay may be performed by first contacting an antibody thathas been immobilized on a solid support, commonly the well of amicrotiter plate, with the sample, such that polypeptides within thesample are allowed to bind to the immobilized antibody. Unbound sampleis then removed from the immobilized polypeptide-antibody complexes anda detection reagent (preferably a second antibody capable of binding toa different site on the polypeptide) containing a reporter group isadded. The amount of detection reagent that remains bound to the solidsupport is then determined using a method appropriate for the specificreporter group.

[0310] More specifically, once the antibody is immobilized on thesupport as described above, the remaining protein binding sites on thesupport are typically blocked. Any suitable blocking agent known tothose of ordinary skill in the art, such as bovine serum albumin orTween 20™ (Sigma Chemical Co., St. Louis, Mo.). The immobilized antibodyis then incubated with the sample, and polypeptide is allowed to bind tothe antibody. The sample may be diluted with a suitable diluent, such asphosphate-buffered saline (PBS) prior to incubation. In general, anappropriate contact time (i.e., incubation time) is a period of timethat is sufficient to detect the presence of polypeptide within a sampleobtained from an individual with breast cancer at least about 95% ofthat achieved at equilibrium between bound and unbound polypeptide.Those of ordinary skill in the art will recognize that the timenecessary to achieve equilibrium may be readily determined by assayingthe level of binding that occurs over a period of time. At roomtemperature, an incubation time of about 30 minutes is generallysufficient.

[0311] Unbound sample may then be removed by washing the solid supportwith an appropriate buffer, such as PBS containing 0.1% Tween 20™. Thesecond antibody, which contains a reporter group, may then be added tothe solid support. Preferred reporter groups include those groupsrecited above.

[0312] The detection reagent is then incubated with the immobilizedantibody-polypeptide complex for an amount of time sufficient to detectthe bound polypeptide. An appropriate amount of time may generally bedetermined by assaying the level of binding that occurs over a period oftime. Unbound detection reagent is then removed and bound detectionreagent is detected using the reporter group. The method employed fordetecting the reporter group depends upon the nature of the reportergroup. For radioactive groups, scintillation counting orautoradiographic methods are generally appropriate. Spectroscopicmethods may be used to detect dyes, luminescent groups and fluorescentgroups. Biotin may be detected using avidin, coupled to a differentreporter group (commonly a radioactive or fluorescent group or anenzyme). Enzyme reporter groups may generally be detected by theaddition of substrate (generally for a specific period of time),followed by spectroscopic or other analysis of the reaction products.

[0313] To determine the presence or absence of a cancer, such as breastcancer, the signal detected from the reporter group that remains boundto the solid support is generally compared to a signal that correspondsto a predetermined cut-off value. In one preferred embodiment, thecut-off value for the detection of a cancer is the average mean signalobtained when the immobilized antibody is incubated with samples frompatients without the cancer. In general, a sample generating a signalthat is three standard deviations above the predetermined cut-off valueis considered positive for the cancer. In an alternate preferredembodiment, the cut-off value is determined using a Receiver OperatorCurve, according to the method of Sackett et al., Clinical Epidemiology:A Basic Science for Clinical Medicine, Little Brown and Co., 1985, p.106-7. Briefly, in this embodiment, the cut-off value may be determinedfrom a plot of pairs of true positive rates (i.e., sensitivity) andfalse positive rates (100%-specificity) that correspond to each possiblecut-off value for the diagnostic test result. The cut-off value on theplot that is the closest to the upper left-hand corner (i.e., the valuethat encloses the largest area) is the most accurate cut-off value, anda sample generating a signal that is higher than the cut-off valuedetermined by this method may be considered positive. Alternatively, thecut-off value may be shifted to the left along the plot, to minimize thefalse positive rate, or to the right, to minimize the false negativerate. In general, a sample generating a signal that is higher than thecut-off value determined by this method is considered positive for acancer.

[0314] In a related embodiment, the assay is performed in a flow-throughor strip test format, wherein the binding agent is immobilized on amembrane, such as nitrocellulose. In the flow-through test, polypeptideswithin the sample bind to the immobilized binding agent as the samplepasses through the membrane. A second, labeled binding agent then bindsto the binding agent-polypeptide complex as a solution containing thesecond binding agent flows through the membrane. The detection of boundsecond binding agent may then be performed as described above. In thestrip test format, one end of the membrane to which binding agent isbound is immersed in a solution containing the sample. The samplemigrates along the membrane through a region containing second bindingagent and to the area of immobilized binding agent. Concentration ofsecond binding agent at the area of immobilized antibody indicates thepresence of a cancer. Typically, the concentration of second bindingagent at that site generates a pattern, such as a line, that can be readvisually. The absence of such a pattern indicates a negative result. Ingeneral, the amount of binding agent immobilized on the membrane isselected to generate a visually discernible pattern when the biologicalsample contains a level of polypeptide that would be sufficient togenerate a positive signal in the two-antibody sandwich assay, in theformat discussed above. Preferred binding agents for use in such assaysare antibodies and antigen-binding fragments thereof. Preferably, theamount of antibody immobilized on the membrane ranges from about 25 ngto about 1 μg, and more preferably from about 50 ng to about 500 ng.Such tests can typically be performed with a very small amount ofbiological sample.

[0315] Of course, numerous other assay protocols exist that are suitablefor use with the tumor proteins or binding agents of the presentinvention. The above descriptions are intended to be exemplary only. Forexample, it will be apparent to those of ordinary skill in the art thatthe above protocols may be readily modified to use tumor polypeptides todetect antibodies that bind to such polypeptides in a biological sample.The detection of such tumor protein specific antibodies may correlatewith the presence of a cancer.

[0316] A cancer may also, or alternatively, be detected based on thepresence of T cells that specifically react with a tumor protein in abiological sample. Within certain methods, a biological samplecomprising CD4⁺ and/or CD8⁺ T cells isolated from a patient is incubatedwith a tumor polypeptide, a polynucleotide encoding such a polypeptideand/or an APC that expresses at least an immunogenic portion of such apolypeptide, and the presence or absence of specific activation of the Tcells is detected. Suitable biological samples include, but are notlimited to, isolated T cells. For example, T cells may be isolated froma patient by routine techniques (such as by Ficoll/Hypaque densitygradient centrifugation of peripheral blood lymphocytes). T cells may beincubated in vitro for 2-9 days (typically 4 days) at 37° C. withpolypeptide (e.g., 5-25 μg/ml). It may be desirable to incubate anotheraliquot of a T cell sample in the absence of tumor polypeptide to serveas a control. For CD4⁺ T cells, activation is preferably detected byevaluating proliferation of the T cells. For CD8⁺ T cells, activation ispreferably detected by evaluating cytolytic activity. A level ofproliferation that is at least two fold greater and/or a level ofcytolytic activity that is at least 20% greater than in disease-freepatients indicates the presence of a cancer in the patient.

[0317] As noted above, a cancer may also, or alternatively, be detectedbased on the level of mRNA encoding a tumor protein in a biologicalsample. For example, at least two oligonucleotide primers may beemployed in a polymerase chain reaction (PCR) based assay to amplify aportion of a tumor cDNA derived from a biological sample, wherein atleast one of the oligonucleotide primers is specific for (i.e.,hybridizes to) a polynucleotide encoding the tumor protein. Theamplified cDNA is then separated and detected using techniques wellknown in the art, such as gel electrophoresis.

[0318] Similarly, oligonucleotide probes that specifically hybridize toa polynucleotide encoding a tumor protein may be used in a hybridizationassay to detect the presence of polynucleotide encoding the tumorprotein in a biological sample.

[0319] To permit hybridization under assay conditions, oligonucleotideprimers and probes should comprise an oligonucleotide sequence that hasat least about 60%, preferably at least about 75% and more preferably atleast about 90%, identity to a portion of a polynucleotide encoding atumor protein of the invention that is at least 10 nucleotides, andpreferably at least 20 nucleotides, in length. Preferably,oligonucleotide primers and/or probes hybridize to a polynucleotideencoding a polypeptide described herein under moderately stringentconditions, as defined above. Oligonucleotide primers and/or probeswhich may be usefully employed in the diagnostic methods describedherein preferably are at least 10-40 nucleotides in length. In apreferred embodiment, the oligonucleotide primers comprise at least 10contiguous nucleotides, more preferably at least 15 contiguousnucleotides, of a DNA molecule having a sequence as disclosed herein.Techniques for both PCR based assays and hybridization assays are wellknown in the art (see, for example, Mullis et al., Cold Spring HarborSymp. Quant. Biol., 51:263, 1987; Erlich ed., PCR Technology, StocktonPress, NY, 1989).

[0320] One preferred assay employs RT-PCR, in which PCR is applied inconjunction with reverse transcription. Typically, RNA is extracted froma biological sample, such as biopsy tissue, and is reverse transcribedto produce cDNA molecules. PCR amplification using at least one specificprimer generates a cDNA molecule, which may be separated and visualizedusing, for example, gel electrophoresis. Amplification may be performedon biological samples taken from a test patient and from an individualwho is not afflicted with a cancer. The amplification reaction may beperformed on several dilutions of cDNA spanning two orders of magnitude.A two-fold or greater increase in expression in several dilutions of thetest patient sample as compared to the same dilutions of thenon-cancerous sample is typically considered positive.

[0321] In another aspect of the present invention, cell capturetechnologies may be used in conjunction, with, for example, real-timePCR to provide a more sensitive tool for detection of metastatic cellsexpressing breast tumor antigens. Detection of breast cancer cells inbiological samples, e.g., bone marrow samples, peripheral blood, andsmall needle aspiration samples is desirable for diagnosis and prognosisin breast cancer patients.

[0322] Immunomagnetic beads coated with specific monoclonal antibodiesto surface cell markers, or tetrameric antibody complexes, may be usedto first enrich or positively select cancer cells in a sample. Variouscommercially available kits may be used, including Dynabeads® EpithelialEnrich (Dynal Biotech, Oslo, Norway), StemSep™ (StemCell Technologies,Inc., Vancouver, BC), and RosetteSep (StemCell Technologies). A skilledartisan will recognize that other methodologies and kits may also beused to enrich or positively select desired cell populations. Dynabeads®Epithelial Enrich contains magnetic beads coated with mAbs specific fortwo glycoprotein membrane antigens expressed on normal and neoplasticepithelial tissues. The coated beads may be added to a sample and thesample then applied to a magnet, thereby capturing the cells bound tothe beads. The unwanted cells are washed away and the magneticallyisolated cells eluted from the beads and used in further analyses.

[0323] RosetteSep can be used to enrich cells directly from a bloodsample and consists of a cocktail of tetrameric antibodies that targetsa variety of unwanted cells and crosslinks them to glycophorin A on redblood cells (RBC) present in the sample, forming rosettes. Whencentrifuged over Ficoll, targeted cells pellet along with the free RBC.The combination of antibodies in the depletion cocktail determines whichcells will be removed and consequently which cells will be recovered.Antibodies that are available include, but are not limited to: CD2, CD3,CD4, CD5, CD8, CD10, CD11b, CD14, CD15, CD16, CD19, CD20, CD24, CD25,CD29, CD33, CD34, CD36, CD38, CD41, CD45, CD45RA, CD45RO, CD56, CD66B,CD66e, HLA-DR, IgE, and TCRαβ.

[0324] Additionally, it is contemplated in the present invention thatmAbs specific for breast tumor antigens can be generated and used in asimilar manner. For example, mAbs that bind to tumor-specific cellsurface antigens may be conjugated to magnetic beads, or formulated in atetrameric antibody complex, and used to enrich or positively selectmetastatic breast tumor cells from a sample. Once a sample is enrichedor positively selected, cells may be lysed and RNA isolated. RNA maythen be subjected to RT-PCR analysis using breast tumor-specific primersin a real-time PCR assay as described herein. One skilled in the artwill recognize that enriched or selected populations of cells may beanalyzed by other methods (e.g., in situ hybridization or flowcytometry).

[0325] In another embodiment, the compositions described herein may beused as markers for the progression of cancer. In this embodiment,assays as described above for the diagnosis of a cancer may be performedover time, and the change in the level of reactive polypeptide(s) orpolynucleotide(s) evaluated. For example, the assays may be performedevery 24-72 hours for a period of 6 months to 1 year, and thereafterperformed as needed. In general, a cancer is progressing in thosepatients in whom the level of polypeptide or polynucleotide detectedincreases over time. In contrast, the cancer is not progressing when thelevel of reactive polypeptide or polynucleotide either remains constantor decreases with time.

[0326] Certain in vivo diagnostic assays may be performed directly on atumor. One such assay involves contacting tumor cells with a bindingagent. The bound binding agent may then be detected directly orindirectly via a reporter group. Such binding agents may also be used inhistological applications. Alternatively, polynucleotide probes may beused within such applications.

[0327] As noted above, to improve sensitivity, multiple tumor proteinmarkers may be assayed within a given sample. It will be apparent thatbinding agents specific for different proteins provided herein may becombined within a single assay. Further, multiple primers or probes maybe used concurrently. The selection of tumor protein markers may bebased on routine experiments to determine combinations that results inoptimal sensitivity. In addition, or alternatively, assays for tumorproteins provided herein may be combined with assays for other knowntumor antigens.

[0328] The present invention further provides kits for use within any ofthe above diagnostic methods. Such kits typically comprise two or morecomponents necessary for performing a diagnostic assay. Components maybe compounds, reagents, containers and/or equipment. For example, onecontainer within a kit may contain a monoclonal antibody or fragmentthereof that specifically binds to a tumor protein. Such antibodies orfragments may be provided attached to a support material, as describedabove. One or more additional containers may enclose elements, such asreagents or buffers, to be used in the assay. Such kits may also, oralternatively, contain a detection reagent as described above thatcontains a reporter group suitable for direct or indirect detection ofantibody binding.

[0329] Alternatively, a kit may be designed to detect the level of mRNAencoding a tumor protein in a biological sample. Such kits generallycomprise at least one oligonucleotide probe or primer, as describedabove, that hybridizes to a polynucleotide encoding a tumor protein.Such an oligonucleotide may be used, for example, within a PCR orhybridization assay. Additional components that may be present withinsuch kits include a second oligonucleotide and/or a diagnostic reagentor container to facilitate the detection of a polynucleotide encoding atumor protein.

[0330] The following Examples are offered by way of illustration and notby way of limitation.

EXAMPLES Example 1 Preparation of Breast Tumor-Specific cDNAs UsingDifferential Display RT-PCR

[0331] This Example illustrates the preparation of cDNA moleculesencoding breast tumor-specific polypeptides using a differential displayscreen.

[0332] A. Preparation of B18Ag1 cDNA and Characterization of mRNAExpression

[0333] Tissue samples were prepared from breast tumor and normal tissueof a patient with breast cancer that was confirmed by pathology afterremoval from the patient. Normal RNA and tumor RNA was extracted fromthe samples and mRNA was isolated and converted into cDNA using a(dT)₁₂AG (SEQ ID NO: 130) anchored 3′ primer. Differential display PCRwas then executed using a randomly chosen primer (CTTCAACCTC) (SEQ IDNO: 103). Amplification conditions were standard buffer containing 1.5mM MgCl₂, 20 pmol of primer, 500 pmol dNTP, and 1 unit of Taq DNApolymerase (Perkin-Elmer, Branchburg, N.J.). Forty cycles ofamplification were performed using 94° C. denaturation for 30 seconds,42° C. annealing for 1 minute, and 72° C. extension for 30 seconds. AnRNA fingerprint containing 76 amplified products was obtained. Althoughthe RNA fingerprint of breast tumor tissue was over 98% identical tothat of the normal breast tissue, a band was repeatedly observed to bespecific to the RNA fingerprint pattern of the tumor. This band was cutout of a silver stained gel, subcloned into the T-vector (Novagen,Madison, Wis.) and sequenced.

[0334] The sequence of the cDNA, referred to as B18Ag1, is provided inSEQ ID NO: 1. A database search of GENBANK and EMBL revealed that theB18Ag1 fragment initially cloned is 77% identical to the endogenoushuman retroviral element S71, which is a truncated retroviral elementhomologous to the Simian Sarcoma Virus (SSV). S71 contains an incompletegag gene, a portion of the pol gene and an LTR-like structure at the 3′terminus (see Werner et al., Virology 174:225-238 (1990)). B18Ag1 isalso 64% identical to SSV in the region corresponding to the P30 (gag)locus. B18Ag1 contains three separate and incomplete reading framescovering a region which shares considerable homology to a wide varietyof gag proteins of retroviruses which infect mammals. In addition, thehomology to S71 is not just within the gag gene, but spans several kb ofsequence including an LTR.

[0335] B18Ag1-specific PCR primers were synthesized using computeranalysis guidelines. RT-PCR amplification (94° C., 30 seconds; 60°C.→42° C., 30 seconds; 72° C., 30 seconds for 40 cycles) confirmed thatB18Ag1 represents an actual mRNA sequence present at relatively highlevels in the patient's breast tumor tissue. The primers used inamplification were B18Ag1-1 (CTG CCT GAG CCA CAA ATG) (SEQ ID NO: 128)and B18Ag1-4 (CCG GAG GAG GAA GCT AGA GGA ATA) (SEQ ID NO: 129) at a 3.5mM magnesium concentration and a pH of 8.5, and B18Ag1-2 (ATG GCT ATTTTC GGG GCC TGA CA) (SEQ ID NO: 126) and B18Ag1-3 (CCG GTA TCT CCT CGTGGG TAT T) (SEQ ID NO: 127) at 2 mM magnesium at pH 9.5. The sameexperiments showed exceedingly low to nonexistent levels of expressionin this patient's normal breast tissue (see FIG. 1). RT-PCR experimentswere then used to show that B18Ag1 mRNA is present in nine other breasttumor samples (from Brazilian and American patients) but absent in, orat exceedingly low levels in, the normal breast tissue corresponding toeach cancer patient. RT-PCR analysis has also shown that the B18Ag1transcript is not present in various normal tissues (including lymphnode, myocardium and liver) and present at relatively low levels in PBMCand lung tissue. The presence of B18Ag1 mRNA in breast tumor samples,and its absence from normal breast tissue, has been confirmed byNorthern blot analysis, as shown in FIG. 2.

[0336] The differential expression of B18Ag1 in breast tumor tissue wasalso confirmed by RNase protection assays. FIG. 3 shows the level ofB18Ag1 mRNA in various tissue types as determined in four differentRNase protection assays. Lanes 1-12 represent various normal breasttissue samples, lanes 13-25 represent various breast tumor samples;lanes 26-27 represent normal prostate samples; lanes 28-29 representprostate tumor samples; lanes 30-32 represent colon tumor samples; lane33 represents normal aorta; lane 34 represents normal small intestine;lane 35 represents normal skin, lane 36 represents normal lymph node;lane 37 represents normal ovary; lane 38 represents normal liver; lane39 represents normal skeletal muscle; lane 40 represents a first normalstomach sample, lane 41 represents a second normal stomach sample; lane42 represents a normal lung; lane 43 represents normal kidney; and lane44 represents normal pancreas. Interexperimental comparison wasfacilitated by including a positive control RNA of known β-actin messageabundance in each assay and normalizing the results of the differentassays with respect to this positive control.

[0337] RT-PCR and Southern Blot analysis has shown the B18Ag1 locus tobe present in human genomic DNA as a single copy endogenous retroviralelement. A genomic clone of approximately 12-18 kb was isolated usingthe initial B18Ag1 sequence as a probe. Four additional subclones werealso isolated by Xbal digestion. Additional retroviral sequencesobtained from the ends of the Xbal digests of these clones (located asshown in FIG. 4) are shown as SEQ ID NO: 3-SEQ ID NO: 10, where SEQ IDNO: 3 shows the location of the sequence labeled 10 in FIG. 4, SEQ IDNO: 4 shows the location of the sequence labeled 11-29, SEQ ID NO: 5shows the location of the sequence labeled 3, SEQ ID NO: 6 shows thelocation of the sequence labeled 6, SEQ ID NO: 7 shows the location ofthe sequence labeled 12, SEQ ID NO: 8 shows the location of the sequencelabeled 13, SEQ ID NO: 9 shows the location of the sequence labeled 14and SEQ ID NO: 10 shows the location of the sequence labeled 11-22.

[0338] Subsequent studies demonstrated that the 12-18 kb genomic clonecontains a retroviral element of about 7.75 kb, as shown in FIGS. 5A and5B. The sequence of this retroviral element is shown in SEQ ID NO: 141.The numbered line at the top of FIG. 5A represents the sense strandsequence of the retroviral genomic clone. The box below this line showsthe position of selected restriction sites. The arrows depict thedifferent overlapping clones used to sequence the retroviral element.The direction of the arrow shows whether the single-pass subclonesequence corresponded to the sense or anti-sense strand. FIG. 5B is aschematic diagram of the retroviral element containing B18Ag1 depictingthe organization of viral genes within the element. The open boxescorrespond to predicted reading frames, starting with a methionine,found throughout the element. Each of the six likely reading frames isshown, as indicated to the left of the boxes, with frames 1-3corresponding to those found on the sense strand.

[0339] Using the cDNA of SEQ ID NO: 1 as a probe, a longer cDNA wasobtained (SEQ ID NO: 227) which contains minor nucleotide differences(less than 1%) compared to the genomic sequence shown in SEQ ID NO: 141.

[0340] B. Preparation of cDNA Molecules Encoding Other BreastTumor-Specific Polypeptides

[0341] Normal RNA and tumor RNA was prepared and mRNA was isolated andconverted into cDNA using a (dT)₁₂AG anchored 3′ primer, as describedabove. Differential display PCR was then executed using the randomlychosen primers of SEQ ID NO: 87-125. Amplification conditions were asnoted above, and bands observed to be specific to the RNA fingerprintpattern of the tumor were cut out of a silver stained gel, subclonedinto either the T-vector (Novagen, Madison, Wis.) or the pCRII vector(Invitrogen, San Diego, Calif.) and sequenced. The sequences areprovided in SEQ ID NO: 11-SEQ ID NO: 86. Of the 79 sequences isolated,67 were found to be novel (SEQ ID NO: 11-26 and 28-77) (see also FIGS.6-20).

[0342] An extended DNA sequence (SEQ ID NO: 290) for the antigen B15Ag1(originally identified partial sequence provided in SEQ ID NO: 27) wasobtained in further studies. Comparison of the sequence of SEQ ID NO:290 with those in the gene bank as described above, revealed homology tothe known human β-A activin gene. Further studies led to the isolationof the full-length cDNA sequence for the antigen B21 GT2 (also referredto as B311D; originally identified partial cDNA sequence provided in SEQID NO: 56). The full-length sequence is provided in SEQ ID NO: 307, withthe corresponding amino acid sequence being provided in SEQ ID NO: 308.Further studies led to the isolation of a splice variant of B311D. TheB311D clone of SEQ ID NO: 316 was sequenced and a XhoI/NotI fragmentfrom this clone was gel purified and 32P-cDTP labeled by random primingfor use as a probe for further screening to obtain additional B311D genesequence. Two fractions of a human breast tumor cDNA bacterial librarywere screened using standard techniques. One of the clones isolated inthis manner yielded additional sequence which includes a poly A+ tail.The determined cDNA sequence of this clone (referred to asB311D_BT1_(—)1A) is provided in SEQ ID NO: 317. The sequences of SEQ IDNO: 316 and 317 were found to share identity over a 464 bp region, withthe sequences diverging near the poly A+ sequence of SEQ ID NO: 317.

[0343] Subsequent studies identified an additional 146 sequences (SEQ IDNO: 142-289), of which 115 appeared to be novel (SEQ ID NO: 142, 143,146-152, 154-166, 168-176, 178-192, 194-198, 200-204, 206, 207, 209-214,216, 218, 219, 221-240, 243-245, 247, 250, 251, 253, 255, 257-266, 268,269, 271-273, 275, 276, 278, 280, 281, 284, 288 and 291). To the best ofthe inventors' knowledge none of the previously identified sequenceshave heretofore been shown to be expressed at a greater level in humanbreast tumor tissue than in normal breast tissue.

[0344] In further studies, several different splice forms of the antigenB11Ag1 (also referred to as B305D) were isolated, with each of thevarious splice forms containing slightly different versions of theB11Ag1 coding frame. Splice junction sequences define individual exonswhich, in various patterns and arrangements, make up the various spliceforms. Primers were designed to examine the expression pattern of eachof the exons using RT-PCR as described below. Each exon was found toshow the same expression pattern as the original B11Ag1 clone, withexpression being breast tumor-, normal prostate- and normaltestis-specific. The determined cDNA sequences for the isolated proteincoding exons are provided in SEQ ID NO: 292-298, respectively. Thepredicted amino acid sequences corresponding to the sequences of SEQ IDNO: 292 and 298 are provided in SEQ ID NO: 299 and 300. Additionalstudies using rapid amplification of cDNA ends (RACE), a 5′ specificprimer to one of the splice forms of B11Ag1 provided above and a breastadenocarcinoma, led to the isolation of three additional, related,splice forms referred to as isoforms B11C-15, B11C-8 and B11C-9, 16. Thedetermined cDNA sequences for these isoforms are provided in SEQ ID NO:301-303, with the corresponding predicted amino acid sequences beingprovided in SEQ ID NO: 304-306.

[0345] The protein coding region of B11C-15 (SEQ ID NO: 301; alsoreferred to as B305D isoform C) was used as a query sequence in a BLASTNsearch of the Genbank DNA database. A match was found to a genomic clonefrom chromosome 21 (Accessson no. AP001465). The pairwise alignmentsprovided in the BLASTN output were used to identify the putative exon,or coding, sequence of the chromosome 21 sequence that corresponds tothe B305D sequence. Based on the BlastN pairwise alignments, thefollowing pieces of GenBank record AP001465 were put together: basepairs 67978-68499, 72870-72987, 73144-73335, 76085-76206, 77905-78085,80520-80624, 87602-87633. This sequence was then aligned with the B305Disoform C sequence using the DNA Star Seqman program and excess sequencewas deleted in such a way as to maintain the sequence most similar toB305D. The final edited form of the chromosome 21 sequence was 96.5%identical to B305D. This resulting edited sequence from chromosome 21was then translated and found to contain no stop codons other than thefinal stop codon in the same position as that for B305D. As with B305D,the chromosome 21 sequence (provided in SEQ ID NO: 325) encoded aprotein (SEQ ID NO: 326) with 384 amino acids. An alignment of thisprotein with the B305D isoform C protein (SEQ ID NO: 304) showed 90%amino acid identity.

[0346] The cDNA sequence of B305D isoform C (SEQ ID NO: 301) was used toidentify homologs by searching the High Throughput Genome Sequencing(HTGS) database (NCBI, National Institutes for Health, Bethesda, Md.).Homologs were identified on Chromosome 2 (Clone ID 9838181), Chromosome10 (Clone ID 10933022), Chromosome 15 (Clone ID 11560284). Thesehomologs shared greater than 90% identity with B305D isoform C at thenucleic acid level. All three of these homologs encode 384 amino acidORFs that share greater than 90% identity with the amino acid sequenceof SEQ ID NO: 304. Further searching of the GenBank database with thesequence of SEQ ID NO: 301 yielded a partial sequence homolog onChromosome 22 (Clone ID 5931507). cDNA sequences for the Chromosome 2,10, 15 and 22 homologs were constructed based on the homology with B305Disoform C and the conserved sequences at intron-exon junctions. The cDNAsequences for the Chromosome 22, 2, 15 and 10 homologs are provided inSEQ ID NO: 327-330, respectively, with the corresponding amino acidsequences being provided in SEQ ID NO: 331, 334, 333 and 332,respectively.

[0347] In subsequent studies on B305D isoform A (cDNA sequence providedin SEQ ID NO: 292), the cDNA sequence (provided in SEQ ID NO: 313) wasfound to contain an additional guanine residue at position 884, leadingto a frameshift in the open reading frame. The determined DNA sequenceof this ORF is provided in SEQ ID NO: 314. This frameshift generates aprotein sequence (provided in SEQ ID NO: 315) of 293 amino acids thatcontains the C-terminal domain common to the other isoforms of B305D butthat differs in the N-terminal region.

Example 2 Preparation of B18AG1 DNA From Human Genomic DNA

[0348] This Example illustrates the preparation of B18Ag1 DNA byamplification from human genomic DNA.

[0349] B18Ag1 DNA may be prepared from 250 ng human genomic DNA using 20pmol of B18Ag1 specific primers, 500 pmol dNTPS and 1 unit of Taq DNApolymerase (Perkin Elmer, Branchburg, N.J.) using the followingamplification parameters: 94° C. for 30 seconds denaturing, 30 seconds60° C. to 42° C. touchdown annealing in 2° C. increments every twocycles and 72° C. extension for 30 seconds. The last increment (a 42° C.annealing temperature) should cycle 25 times. Primers were selectedusing computer analysis. Primers synthesized were B18Ag1-1, B18Ag1-2,B18Ag1-3, and B18Ag1-4. Primer pairs that may be used are 1+3, 1+4, 2+3,and 2+4.

[0350] Following gel electrophoresis, the band corresponding to B18Ag1DNA may be excised and cloned into a suitable vector.

Example 3 Preparation of B18Ag1 DNA From Beast Tumor cDNA

[0351] This Example illustrates the preparation of B18Ag1 DNA byamplification from human breast tumor cDNA.

[0352] First strand cDNA is synthesized from RNA prepared from humanbreast tumor tissue in a reaction mixture containing 500 ng poly A+ RNA,200 pmol of the primer (T)₁₂AG (i.e., TTT TTT TTT TTT AG) (SEQ ID NO:130), 1×first strand reverse transcriptase buffer, 6.7 mM DTT, 500 mmoldNTPs, and 1 unit AMV or MMLV reverse transcriptase (from any supplier,such as Gibco-BRL (Grand Island, N.Y.)) in a final volume of 30 μl.After first strand synthesis, the cDNA is diluted approximately 25 foldand 1 μl is used for amplification as described in Example 2. While someprimer pairs can result in a heterogeneous population of transcripts,the primers B18Ag1-2 (5′ATG GCT ATT TTC GGG GGC TGA CA) (SEQ ID NO: 126)and B18Ag1-3 (5′CCG GTA TCT CCT CGT GGG TAT T) (SEQ ID NO: 127) yield asingle 151 bp amplification product.

Example 4 Identification of B-cell and T-cell Epitopes of B18AG1

[0353] This Example illustrates the identification of B18Ag1 epitopes.

[0354] The B18Ag1 sequence can be screened using a variety of computeralgorithms. To determine B-cell epitopes, the sequence can be screenedfor hydrophobicity and hydrophilicity values using the method of Hopp,Prog. Clin. Biol. Res. 172B:367-77 (1985) or, alternatively, Cease etal., J. Exp. Med. 164:1779-84 (1986) or Spouge et al., J. Immunol.138:204-12 (1987). Additional Class II MHC (antibody or B-cell) epitopescan be predicted using programs such as AMPHI (e.g., Margalit et al., J.Immunol. 138:2213 (1987)) or the methods of Rothbard and Taylor (e.g.,EMBO J. 7:93 (1988)).

[0355] Once peptides (15-20 amino acids long) are identified using thesetechniques, individual peptides can be synthesized using automatedpeptide synthesis equipment (available from manufacturers such as PerkinElmer/Applied Biosystems Division, Foster City, Calif.) and techniquessuch as Merrifield synthesis. Following synthesis, the peptides can usedto screen sera harvested from either normal or breast cancer patients todetermine whether patients with breast cancer possess antibodiesreactive with the peptides. Presence of such antibodies in breast cancerpatient would confirm the immunogenicity of the specific B-cell epitopein question. The peptides can also be tested for their ability togenerate a serologic or humoral immune in animals (mice, rats, rabbits,chimps etc.) following immunization in vivo. Generation of apeptide-specific antiserum following such immunization further confirmsthe immunogenicity of the specific B-cell epitope in question.

[0356] To identify T-cell epitopes, the B18Ag1 sequence can be screenedusing different computer algorithms which are useful in identifying 8-10amino acid motifs within the B18Ag1 sequence which are capable ofbinding to HLA Class I MHC molecules. (see, e.g., Rammensee et al.,Immunogenetics 41:178-228 (1995)). Following synthesis such peptides canbe tested for their ability to bind to class I MHC using standardbinding assays (e.g., Sette et al., J. Immunol. 153:5586-92 (1994)) andmore importantly can be tested for their ability to generate antigenreactive cytotoxic T-cells following in vitro stimulation of patient ornormal peripheral mononuclear cells using, for example, the methods ofBakker et al., Cancer Res. 55:5330-34 (1995); Visseren et al., J.Immunol. 154:3991-98 (1995); Kawakami et al., J. Immunol. 154:3961-68(1995); and Kast et al., J. Immunol. 152:3904-12 (1994). Successful invitro generation of T-cells capable of killing autologous (bearing thesame Class I MHC molecules) tumor cells following in vitro peptidestimulation further confirms the immunogenicity of the B18Ag1 antigen.Furthermore, such peptides may be used to generate murine peptide andB18Ag1 reactive cytotoxic T-cells following in vivo immunization in micerendered transgenic for expression of a particular human MHC Class Ihaplotype (Vitiello et al., J. Exp. Med. 173:1007-15 (1991).

[0357] A representative list of predicted B18Ag1 B-cell and T-cellepitopes, broken down according to predicted HLA Class I MHC bindingantigen, is shown below:

[0358] Predicted Th Motifs (B-cell epitopes) (SEQ ID NOS.: 131-133)

[0359] SSGGRTFDDFHRYLLVGI

[0360] QGAAQKPINLSKXIEVVQGHDE

[0361] SPGVFLEHLQEAYRIYTPFDLSA

[0362] Predicted HLA A2.1 Motifs (T-cell epitopes) (SEQ ID NOS.:134-140)

[0363] YLLVGIQGA

[0364] GAAQKPINL

[0365] NLSKXIEVV

[0366] EVVQGHDES

[0367] HLQEAYRIY

[0368] NLAFVAQAA

[0369] FVAQAAPDS

Example 5 Identification of T-cell Epitopes of B11AG1

[0370] This Example illustrates the identification of B11Ag1 (alsoreferred to as B305D) epitopes. Four peptides, referred to as B11-8,B11-1, B11-5 and B11-12 (SEQ ID NO: 309-312, respectfully) were derivedfrom the B11Ag1 gene.

[0371] Human CD8 T cells were primed in vitro to the peptide B11-8 usingdendritic cells according to the protocol of Van Tsai et al. (CriticalReviews in Immunology 18:65-75, 1998). The resulting CD8 T cell cultureswere tested for their ability to recognize the B11-8 peptide or anegative control peptide, presented by the B-LCL line, JY. Briefly, Tcells were incubated with autologous monocytes in the presence of 10ug/ml peptide, 10 ng/ml IL-7 and 10 ug/ml IL-2, and assayed for theirability to specifically lyse target cells in a standard 51-Cr releaseassay. As shown in FIG. 22, the bulk culture line demonstrated strongrecognition of the B11-8 peptide with weaker recognition of the peptideB11-1.

[0372] A clone from this CTL line was isolated following rapid expansionusing the monoclonal antibody OKT3 and human IL-2. As shown in FIG. 23,this clone (referred to as A1), in addition to being able to recognizespecific peptide, recognized JY LCL transduced with the B11Ag1 gene.This data demonstrates that B11-8 is a naturally processed epitope ofthe B11Ag1 gene. In addition these T cells were further found torecognize and lyse, in an HLA-A2 restricted manner, an established tumorcell line naturally expressing B11Ag1 (FIG. 24). The T cells stronglyrecognize a lung adenocarcinoma (LT-140-22) naturally expressing B11Ag1transduced with HLA-A2, as well as an A2+ breast carcinoma (CAMA-1)transduced with B11Ag1, but not untransduced lines or another negativetumor line (SW620).

[0373] These data clearly demonstrate that these human T cells recognizenot only B11-specific peptides but also transduced cells, as well asnaturally expressing tumor lines.

[0374] CTL lines raised against the antigens B11-5 and B11-12, using theprocedures described above, were found to recognize correspondingpeptide-coated targets.

Example 6 Characterization of Breast Tumor Genes Discovered byDifferential Display PCR

[0375] The specificity and sensitivity of the breast tumor genesdiscovered by differential display PCR were determined using RT-PCR.This procedure enabled the rapid evaluation of breast tumor gene mRNAexpression semiquantitatively without using large amounts of RNA. Usinggene specific primers, mRNA expression levels in a variety of tissueswere examined, including 8 breast tumors, 5 normal breasts, 2 prostatetumors, 2 colon tumors, 1 lung tumor, and 14 other normal adult humantissues, including normal prostate, colon, kidney, liver, lung, ovary,pancreas, skeletal muscle, skin, stomach and testes.

[0376] To ensure the semiquantitative nature of the RT-PCR, β-actin wasused as internal control for each of the tissues examined. Serialdilutions of the first strand cDNAs were prepared and RT-PCR assaysperformed using β-actin specific primers. A dilution was then selectedthat enabled the linear range amplification of β-actin template, andwhich was sensitive enough to reflect the difference in the initial copynumber. Using this condition, the β-actin levels were determined foreach reverse transcription reaction from each tissue. DNA contaminationwas minimized by DNase treatment and by assuring a negative result whenusing first strand cDNA that was prepared without adding reversetranscriptase.

[0377] Using gene specific primers, the mRNA expression levels weredetermined in a variety of tissues. To date, 38 genes have beensuccessfully examined by RT-PCR, five of which exhibit good specificityand sensitivity for breast tumors (B15AG-1, B31GA1b, B38GA2a, B11A1a andB18AG1a). FIGS. 21A and 21B depict the results for three of these genes:B15AG-1 (SEQ ID NO: 27), B31GA1b (SEQ ID NO: 148) and B38GA2a (SEQ IDNO: 157). Table 2 summarizes the expression level of all the genestested in normal breast tissue and breast tumors, and also in othertissues. TABLE 2 Percentage of Breast Cancer Antigens That Are ExpressedIn Various Tissues Breast Tissues Over-expressed in Breast Tumors 84%Equally Expressed in Normals and Tumor 16% Other Tissues Over-expressedin Breast Tumors  9% but not in any Normal Tissues Over-expressed inBreast Tumors 30% but Expressed in Some Normal Tissues Over-expressed inBreast Tumors 61% but Equally Expressed in All Other Tissues

Example 7 Preparation and Characterization of Antibodies Against BreastTumor Polypeptides

[0378] Polyclonal antibodies against the breast tumor antigen B305D wereprepared as follows.

[0379] The breast tumor antigen expressed in an E. coli recombinantexpression system was grown overnight in LB broth with the appropriateantibiotics at 37° C. in a shaking incubator. The next morning, 10 ml ofthe overnight culture was added to 500 ml to 2×YT plus appropriateantibiotics in a 2L-baffled Erlenmeyer flask. When the Optical Density(at 560 nm) of the culture reached 0.4-0.6, the cells were induced withIPTG (1 mM). Four hours after induction with IPTG, the cells wereharvested by centrifugation. The cells were then washed with phosphatebuffered saline and centrifuged again. The supernatant was discarded andthe cells were either frozen for future use or immediately processed.Twenty ml of lysis buffer was added to the cell pellets and vortexed. Tobreak open the E. coli cells, this mixture was then run through theFrench Press at a pressure of 16,000 psi. The cells were thencentrifuged again and the supernatant and pellet were checked bySDS-PAGE for the partitioning of the recombinant protein. For proteinsthat localized to the cell pellet, the pellet was resuspended in 10 mMTris pH 8.0, 1% CHAPS and the inclusion body pellet was washed andcentrifuged again. This procedure was repeated twice more. The washedinclusion body pellet was solubilized with either 8 M urea or 6 Mguanidine HCl containing 10 mM Tris pH 8.0 plus 10 mM imidazole. Thesolubilized protein was added to 5 ml of nickel-chelate resin (Qiagen)and incubated for 45 min to 1 hour at room temperature with continuousagitation. After incubation, the resin and protein mixture were pouredthrough a disposable column and the flow through was collected. Thecolumn was then washed with 10-20 column volumes of the solubilizationbuffer. The antigen was then eluted from the column using 8M urea, 10 mMTris pH 8.0 and 300 mM imidazole and collected in 3 ml fractions. ASDS-PAGE gel was run to determine which fractions to pool for furtherpurification.

[0380] As a final purification step, a strong anion exchange resin suchas HiPrepQ (Biorad) was equilibrated with the appropriate buffer and thepooled fractions from above were loaded onto the column. Antigen waseluted off the column with a increasing salt gradient. Fractions werecollected as the column was run and another SDS-PAGE gel was run todetermine which fractions from the column to pool. The pooled fractionswere dialyzed against 10 mM Tris pH 8.0. The protein was then vialedafter filtration through a 0.22 micron filter and the antigens werefrozen until needed for immunization.

[0381] Four hundred micrograms of B305D antigen was combined with 100micrograms of muramyldipeptide (MDP). Every four weeks rabbits wereboosted with 100 micrograms mixed with an equal volume of IncompleteFreund's Adjuvant (IFA). Seven days following each boost, the animal wasbled. Sera was generated by incubating the blood at 4° C. for 12-24hours followed by centrifugation.

[0382] Ninety-six well plates were coated with B305D antigen byincubating with 50 microliters (typically 1 microgram) of recombinantprotein at 4° C. for 20 hours. 250 microliters of BSA blocking bufferwas added to the wells and incubated at room temperature for 2 hours.Plates were washed 6 times with PBS/0.01% Tween. Rabbit sera was dilutedin PBS. Fifty microliters of diluted sera was added to each well andincubated at room temperature for 30 min. Plates were washed asdescribed above before 50 microliters of goat anti-rabbit horse radishperoxidase (HRP) at a 1:10000 dilution was added and incubated at roomtemperature for 30 min. Plates were again washed as described above and100 microliters of TMB microwell peroxidase substrate was added to eachwell. Following a 15 min incubation in the dark at room temperature, thecolorimetric reaction was stopped with 100 microliters of 1N H₂SO₄ andread immediately at 450 nm. The polyclonal antibodies showedimmunoreactivity to B305D.

[0383] Immunohistochemical (IHC) analysis of B305D expression in breastcancer and normal breast specimens was performed as follows.Paraffin-embedded formal fixed tissue was sliced into 8 micron sections.Steam heat induced epitope retrieval (SHIER) in 0.1 M sodium citratebuffer (pH 6.0) was used for optimal staining conditions. Sections wereincubated with 10% serum/PBS for 5 minutes. Primary antibody was addedto each section for 25 min at indicated concentrations followed by a 25min incubation with either an anti-rabbit or anti-mouse biotinylatedantibody. Endogenous peroxidase activity was blocked by three 1.5 minincubations with hydrogen peroxide. The avidin biotincomplex/horseradish peroxidase (ABC/HRP) systems was used along with DABchromagen to visualize antigen expression. Slides were counterstainedwith hematoxylin. B305D expression was detected in both breast tumor andnormal breast tissue. However, the intensity of staining was much lessin normal samples than in tumor samples and surface expression of B305Dwas observed only in breast tumor tissues.

[0384] A summary of real-time PCR and immunohistochemical analysis ofB305D expression in an extensive panel of normal tissues is presented inTable 3 below. These results demonstrate minimal expression of B305D intestis, inconclusive results in gall bladder, and no detection in allother tissues tested. TABLE 3 mRNA IHC staining Tissue type SummaryModerately Positive Testis Nuclear staining of positive small minorityof spermatids; spermatozoa negative; siminoma negative Negative NegativeThymus No expression N/A Negative Artery No expression Negative NegativeSkeletal muscle No expression Negative Positive (weak Small bowel Noexpression staining) Negative Positive (weak Ovary No expressionstaining) Negative Pituitary No expression Negative Positive (weakStomach No expression staining) Negative Negative Spinal cord Noexpression Negative Negative Spleen No expression Negative NegativeUreter No expression N/A Negative Gall bladder Inconclusive N/A NegativePlacenta No expression Negative Negative Thyroid No expression NegativeNegative Heart No expression Negative Negative Kidney No expressionNegative Negative Liver No expression Negative Negative Brain- Noexpression cerebellum Negative Negative Colon No expression NegativeNegative Skin No expression Negative Negative Bone marrow No expressionN/A Negative Parathyroid No expression Negative Negative Lung Noexpression Negative Negative Esophagus No expression Negative Positive(weak Uterus No expression staining) Negative Negative Adrenal Noexpression Negative Negative Pancreas No expression N/A Negative Lymphnode No expression Negative Negative Brain-cortex No expression N/ANegative Fallopian tube No expression Negative Positive (weak Bladder Noexpression staining) Negative N/A Bone No expression Negative N/ASalivary gland No expression Negative N/A Activated No expression PBMCNegative N/A Resting PBMC No expression Negative N/A Trachea Noexpression Negative N/A Vena cava No expression Negative N/A Retina Noexpression Negative N/A Cartilage No expression

Example 8 Protein Expression of Breast Tumor Antigens

[0385] This example describes the expression and purification of thebreast tumor antigen B305D in E. coli and in mammalian cells.

[0386] Expression of B305D isoform C-15 (SEQ ID NO: 301; translated to384 amino acids) in E. coli was achieved by cloning the open readingframe of B305D isoform C-15 downstream of the first 30 amino acids ofthe M. tuberculosis antigen Ra12 (SEQ ID NO: 318) in pET17b. First, theinternal EcoRI site in the B305D ORF was mutated without changing theprotein sequence so that the gene could be cloned at the EcoRI site withRa12. The PCR primers used for site-directed mutagenesis are shown inSEQ ID NO: 319 (referred to as AW012) and SEQ ID NO: 320 (referred to asAW013). The ORF of EcoRI site-modified B305D was then amplified by PCRusing the primers AW014 (SEQ ID NO: 321) and AW015 (SEQ ID NO: 322). ThePCR product was digested with EcoRI and ligated to the Ra12/pET17bvector at the EcoRI site. The sequence of the resulting fusion construct(referred to as Ra12mB11C) was confirmed by DNA sequencing. Thedetermined cDNA sequence for the fusion construct is provided in SEQ IDNO: 323, with the amino acid sequence being provided in SEQ ID NO: 324.

[0387] The fusion construct was transformed into BL21(DE3)CodonPlus-RILE. coli (Stratagene) and grown overnight in LB broth with kanamycin. Theresulting culture was induced with IPTG. Protein was transferred to PVDFmembrane and blocked with 5% non-fat milk (in PBS-Tween buffer), washedthree times and incubated with mouse anti-His tag antibody (Clontech)for 1 hour. The membrane was washed 3 times and probed with HRP-ProteinA (Zymed) for 30 min. Finally, the membrane was washed 3 times anddeveloped with ECL (Amersham). Expression was detected by Western blot.

[0388] For recombinant expression in mammalian cells, B305D isoform C-15(SEQ ID NO: 301; translated to 384 amino acids) was subcloned into themammalian expression vectors pCEP4 and pcDNA3.1 (Invitrogen). Theseconstructs were transfected into HEK293 cells (ATCC) using Fugene 6reagent (Roche). Briefly, the HEK cells were plated at a density of100,000 cells/ml in DMEM (Gibco) containing 10% FBS (Hyclone) and grownovernight. The following day, 2 ul of Fugene 6 was added to 100 ul ofDMEM containing no FBS and incubated for 15 minutes at room temperature.The Fugene 6/DMEM mixture was added to 1 ug of B305D/pCEP4 orB305D/pcDNA plasmid DNA and incubated for 15 minutes at roomtemperature. The Fugene/DNA mix was then added to the HEK293 cells andincubated for 48-72 hours at 37° C. with 7% CO₂. Cells were rinsed withPBS, the collected and pelleted by centrifugation.

[0389] For Western blot analysis, whole cell lysates were generated byincubating the cells in Triton-X100 containing lysis buffer for 30minutes on ice. Lysates were then cleared by centrifugation at 10,000rpm for 5 minutes at 4° C. Samples were diluted with SDS-PAGE loadingbuffer containing beta-mercaptoethanol, and boiled for 10 minutes priorto loading the SDS-PAGE gel. Proteins were transferred to nitrocelluloseand probed using Protein A purified anti-B305D rabbit polyclonal sera(prepared as described above) at a concentration of 1 ug/ml. The blotwas revealed with a goat anti-rabbit lg coupled to HRP followed byincubation in ECL substrate. Expression of B305D was detected in theHEK293 lysates transfected with B305D, but not in control HEK293 cellstransfected with vector alone.

[0390] For FACS analysis, cells were washed further with ice coldstaining buffer and then incubated with a 1:100 dilution of a goatanti-rabbit lg (H+L)-FITC reagent (Southern Biotechnology) for 30minutes on ice. Following 3 washes, the cells were resuspended instaining buffer containing Propidium Iodide (PI), a vital stain thatallows for identification of permeable cells, and then analyzed by FACS.The FACS analysis showed surface expression of B305D protein.

Example 9 Expresion of Full-Length B305D in Insect Cells Using aBaculovirus Expression System

[0391] The cDNA for the full-length breast tumor antigen, B305D isoformC (SEQ ID NO: 301), with a C-terminal His Tag was made by PCR usingB11C15/pBib as a template and the following primers:

[0392] B305DF1 (SEQ ID NO: 337):

[0393] 5′CGGCGGATCCACCATGGTGGTTGAGGTTGATTCC

[0394] B305DRV1 (SEQ ID NO: 338):

[0395]5′CGGCTCTAGATTAATGGTGATGGTGATGATGATGGTGATGATGTTTATTTCTGGTTCTTGAGACATTTTCTGGA.

[0396] The PCR product with the expected size was recovered from anagarose gel, digested with the Bam Hi and Xba I restriction enzymes, andligated into the transfer plasmid pFastBac1 which was digested with thesame restriction enzymes. The sequence of the insert was confirmed byDNA sequencing and is set forth in SEQ ID NO: 335. The predicted aminoacid sequence of B305D with the C-terminal His tag is set forth in SEQID NO: 336. The recombinant transfer plasmid pFBB305D was used to makerecombinant bacmid DNA and virus by the Bac-To-Bac baculovirusexpression system (Invitrogen Life Technologies, Carlsbad, Calif.). Therecombinant BVB305D virus was amplified in Sf9 insect cells and used toinfect High Five insect cells. Infected cells were harvested at 24-30hours post-infection. The identity of the recombinant protein wasconfirmed by Western blot with a rabbit polyclonal antibody againstB305D. Recombinant protein was further analyzed by SDS-PAGE followed byCoomassie blue staining.

Example 10 Identification of an Additional B305D Homolog discovered byBioinformatic Search

[0397] The High Throughput Genome Sequencing (HTGS) database wassearched with the B305D C form sequence (SEQ ID NO: 301) and revealedanother highly related copy of the B305D gene, tentatively localized toChromosome 14. The sequences identified were spliced together based onthe B305D C form sequence and exon-intron splice sites. This predictedcDNA sequence (SEQ ID NO: 339) was translated to generate the predictedamino acid sequence (SEQ ID NO: 340). The B305D gene family members havebeen shown to be overexpressed in breast cancer, prostate cancer, andovarian cancer.

Example 11 Immunohistochemical (IHC) Analysis of B305D Expression

[0398] Analysis suggests that B305D is a type II plasma membrane proteinof about 43 kDa with 1 predicted trasmembrane spanning domain. There areno glycosylation sites and its function remains unknown. Disclosedherein is further examination of B305D expression byimmunohistochemistry (IHC) analysis in a variety of tumor and normaltissues.

[0399] Methods and Materials:

[0400] In order to determine which tissues express the breast cancerantigen B305D, IHC analysis was performed on a diverse range of tissuesections. Tissue samples were fixed in formalin solution for 12-24 hoursand embedded in paraffin before being sliced into 8 micron sections.Steam heat induced epitope retrieval (SHIER) in 0.1 M sodium citratebuffer (pH 6.0) was used for optimal staining conditions. Sections wereincubated with 10% serum/PBS for 5 minutes. Primary antibody was addedto each section for 25 minutes at indicated concentrations followed by25 minute incubation with anti-rabbit biotinylated antibody. Endogenousperoxidase activity was blocked by three 1.5 minute incubations withhydrogen peroxidase. The avidin biotin complex/horse radish peroxidase(ABC/HRP) system was used along with DAB chromogen to visualize antigenexpression. Slides were counterstained with hematoxylin to visualizecell nuclei.

[0401] Rabbit polyclonal antibodies against B305D were shown in Example7 to react in formalin fixed, paraffin-embedded tissues. The antibodywas shown to label the plasma membrane of a subset of breast carcinomas.B305D was shown to label tissues that were positive for cerb-2, alsocalled Her-2/neu. HER-2/neu (p185) is the protein product of theHER-2/neu oncogene. The HER-2/neu gene is amplified and the HER-2/neuprotein is overexpressed in a variety of cancers including breast,ovarian, colon, lung, prostate and hematological cancers. HER-2/neu isrelated to malignant transformation and is found in 50%-60% of ductal insitu carcinoma and 20%-40% of all breast cancers, as well as asubstantial fraction of adenocarcinomas arising in the ovaries,prostate, colon and lung. HER-2/neu is intimately associated not onlywith the malignant phenotype, but also with the aggressiveness of themalignancy, being found in one-fourth of all invasive breast cancers.HER-2/neu overexpression is correlated with a poor prognosis in bothbreast and ovarian cancer. In this study breast carcinomas were testedfrom two age groups; women under 50 at the time of tumor removal andwomen over 50 at the time of tumor removal. B305D staining was evaluatedfor each. In addition to breast carcinomas ovarian carcinomas, normalpancreas, normal kidney and normal stomach were tested for B305Dreactivity.

[0402] Formalin-fixed, paraffin-embedded breast carcinomas from 23different patients were tested for B305D reactivity. The age of thepatient at the time of tumor removal was available in all cases todetermine whether patient age is associated with B305D staining. In manycases, estrogen receptor/progesterone receptor (ER/PR) data and cerb2data was available from the pathology reports. Breast patients werechosen simply based on age. These patients in the ‘younger’ group areclose to the age of 40. We also obtained tumors from patients that werecloser to the age of 70. This group is referred to as the ‘older’ group.

[0403] In addition to breast carcinomas, 17 different ovarian carcinomaswere immunohistochemically analyzed for B305D staining. Five sampleseach of normal stomach, kidney and pancreas were also tested. For mostof the tissues, the B305D antibody was tested with two differentdetection systems, one with ABC as the Horseradish Peroxidase (HRP)enzyme-linked reagent and another with strept-avidin as the HRP reagent.In all cases, rabbit IgG was run as a negative control in parallel withthe B305D antibody. B305D was tested at 2.5 μg/ml using SHIER II heatpretreatment. Breast carcinoma multi-tissue block, QMTB21, was used as apositive control for the antibody. Tumor #5 in the block was previouslyshown to label with a membrane pattern with the B305D antibody.

[0404] Results: Breast Carcinomas (Results Shown in Table 4)

[0405] The avidin-biotin complex (ABC) stained slides were lighter thanexpected, although membrane staining was detected in the positivecontrol. To make sure that no positive staining was overlooked, theslides were tested with the strept-avidin (SA) detection. Upon theanalysis of the ABC slides, only one tumor labeled with a membranepattern. This tumor was from a 42 yr old patient who also demonstratedmembrane staining for cerb2. When retested with SA, an older patientthat was cerb2 membrane positive was included. This tumor was from an 80yr old patient. Breast cancer staining results are outlined in Table 4below. The staining data presented in tables 4-6 is from the SA-HRPstaining. The B305D antibody labels breast carcinomas in the cytoplasmand on the plasma membrane. Membrane staining is limited to tumor cells,whereas cytoplasmic staining is also often present in the normal ductalepithelium. Among the SA labeled tissues, only the positive control andthe 42 yr old and the 80 yr old that were cerb2 positive labeledmembrane positive for B305D. Two other cases labeled with light membranestaining in a minority of tumor cells. One case was from a 28 yr oldpatient, the other from a 73 yr old patient; cerb2 status was notavailable for either of these cases. The limited staining in these twocases with lighter staining may be due to tissue fixation as positivecells were found on the periphery of the tissue.

[0406] Thus, 4 cases of 23 (less than 20%) labeled with a membranepattern for B305D. Less than 10% of the tumors (2 of 23) labeled withdefinitive membrane staining. In a previous random study, 3 of 15 casesdemonstrated membrane staining for B305D. Cerb2 data was not availablefor all of the tissues tested but for the two cases that weredefinitively positive for B305D, both were strongly positive for cerb2.B305D membrane positive cases were split evenly across the ‘younger’ and‘older’ groups. The younger group included 11 patients under 50 and theolder group included 12 patients 50 or older. Of this older group, 9 ofthe patients are 66 or older, and 7 were in their 70's and 80's (onetumor from a 50 year old had only a small amount of tumor in the blockand may be discounted—thus 4 of 22 positive). ER/PR data was availablefor most cases but no association with B305D could be determined. Thus,based on this and previous IHC data, B305D expression is closelyassociated with cerb2 expression. Further B305D testing of cerb2positive breast tumors may strengthen this correlation. From the resultsof this study, patient age at the time of tumor 10 removal does notappear to correlate with B305D staining. TABLE 4 AGE RELATED B305DREACTIVITY IN BREAST CARCINOMAS B305D IHC Accession No. Age ReactivityDiagnosis ER/PR Status S86-2763 29 Cytoplasmic staining InfiltratingDuctal ER/PR negative (slide 1) S00-9327 28 Marginal membraneInfiltrating N/A (slide 2) staining Lobular S00-4786 43 Lightcytoplasmic Infiltrating Mixed ER positive 2-3+ (slide 3) stainingDuctal/Lobular PR positive 2-3+ Cerb2 Negative 1+ S86-1877 40Cytoplasmic staining Infiltrating Ductal ER positive (slide 4) PRstrongly positive S84-2015 40 Light cytoplasmic Infiltrating Ductal N/A(slide 5) staining S88-1981 40 Cytoplasmic staining Infiltrating DuctalN/A (slide 6) S84-2915 38 Light cytoplasmic Infiltrating Ductal ERstrongly (slide 7) staining positive PR positive S86-1510 41Infiltrating Ductal ER positive (slide 8) PR strongly positive S01-31(slide 9) 42 Membrane staining; Infiltrating Ductal Cerb2 positive 3+cytoplasmic staining S84-855 48 Light cytoplasmic Infiltrating ducal ERPositive (slide 10) staining PR strongly positive 00-1826 46 Lightcytoplasmic Infiltrating ducal ER-positive 3+ (slide 50) stainingPR-positive 3+ S00-2297 50 Light cytoplasmic Infiltrating ducalER-negative (slide 11) staining PR-positive 1+ Cerb2 negative 1+S00-3232A 50 Light cytoplasmic Infiltrating ducal ER-positive 3+ (slide12) staining (very little PR-positive 3+ tumor) Cerb2-negative 1+S00-8096 54 Infiltrating ducal ER-Negative (slide 13) PR-NegativeCerb2-negative 1+ S00-2097 66 Very little tumor Infiltrating ducalER-positive 3+ (slide 14) PR-positive 2-3+ Cerb2-negative 2+ S88-2476 79Infiltrating ducal ER-strongly (slide 15) positive PR-strongly positiveS88-2551 81 Very light Infiltrating ducal ER-strongly (slide 16)cytoplasmic staining positive PR-positive S88-2665 73 Marginal membraneInfiltrating ducal ER-positive (slide 17) staining; cytoplasmicPR-negative staining S88-2476 79 Light membrane Infiltrating ducalER-strongly (slide 18) staining positive PR-strongly positive S00-249177 Light cytoplasmic Lobular ER-positive 1-3+ (slide 19) stainingInfiltrating PR-positive 1-3+ Little tumor present Cerb2-negative 3+S85-2667 68 Cytoplasmic staining Infiltrating ducal ER-strongly (slide20) positive PR-strongly positive 00-6606A 80 Membrane staining;Infiltrating ducal ER-negative (slide 49) cytoplasmic stainingPR-negative Cerb2-positive 3+ S88-1146 88 Light cytoplasmic Infiltratingducal ER-strongly (slide 50, in staining positive box 1) PR-negative

[0407] Ovarian Carcinomas (Results Outlined in Table 5)

[0408] None of the 17 ovarian carcinomas tested with the B305D antibodylabeled with a membrane pattern. About half of the tissues labeled witha cytoplasmic staining pattern. TABLE 5 B305D STAINING OF OVARIANCARCINOMAS IHC Tissue (slide #) Age Diagnosis Reactivity/Comments 1.73-1808 (slide 73 Papillary mucinous 37) adenocarcinoma 2. 76-1076(slide 50 Serous adenocarcinoma 38) 3. 81-1910 (slide 51 Serousadenocarcinoma Cytoplasmic staining; not 39) uniform 4. 88-220 (slide40) 40 Mucinous Light cytoplasmic staining cystadenocarcinoma 5. 88-2207(slide 75 Papillary Serious 41) cystadenocarcinoma 6. 88-2527 (slide 29Malignant teratoma Light cytoplasmic 42) staining; not uniform 7.00-5294 (slide 55 Papillary adenocarcinoma Light cytoplasmic staining43) 8. 84-779 (slide 44) 48 Endometriod carcinoma Light cytoplasmicstaining 9. 84-1843 (slide 32 Papillary serious Cytoplasmic staining 45)adenocarcinoma 10. 85-2373 (slide 47 Granulosa cell tumor Lightcytoplasmic staining 46) 11. 86-813 (slide 47) 74 Clear cell carcinoma12. QMTB#26 (slide Five different ovarian All negative 48) carcinomas

[0409] Normal Tissues (Results Outlined in Table6)

[0410] Of the five stomach cases tested, all had staining abovebackground in the glands below the gastric epithelium. Staining wascytoplasmic and grainy and was present with both detection systems.There was some staining in the negative control but this staining wasdiffuse and not grainy. Background staining was common in these cells.The B305D staining appeared to be due to the antibody binding and notthe detection system.

[0411] Five different kidney cases were tested. The medulla region wasrepresented in each case. There was staining in the tubules throughoutthe kidney, but this appears to be due to endogenous biotin as similarbut lighter staining was present in the negative controls. There wasmuch less staining in the ABC stained slides compared with thestrept-avidin slides, which is also consistent with endogenous biotin.The SHIER II pretreatment required to obtain staining with the antibodytended to give more background staining, particularly due to endogenousbiotin.

[0412] Of the five different pancreas tissues tested, no specificstaining was detected. A subset of acinar cells gave staining in boththe B305D and the rabbit IgG control. Once again this staining wasnon-specific. Pancreas often gave non-specific staining, possibly due tothe enzymatic activity of the tissue.

[0413] A variety of other normal tissues (not shown in Table 6) weretested including skin, testis, colon, heart, thymus, artery, skeletalmuscle, small bowel, pituitary, spinal cord, spleen, ureter, gallbladder, placenta, thyroid, liver, brain-cerebellum, bone marrow,parathyroid, lung esophagus, uterus, adrenal, lymph node, brain-cortex,fallopian tude, bladder, and prostate. Weak IHC staining was observed insmall bowel, uterus, and bladder. However, no mRNA expression was seenin these tissues. Thus, this weak staining likely does not representprotein expression in these tissues. The gall bladder stained positiveand will be analyzed further. Half of the prostate samples stainedpositive as well as the single testis sample examined.

[0414] B305D expression was also analyzed in prostate tumor samples. Oneof 5 grade 3+3 samples stained positive while none of the grade 3+4samples stained positive. One additional sample of 3 unknown gradesamples stained positive. However, an additional array of 55 primary andprimary metastatic prostate tumor samples was tested and no staining wasobserved. TABLE 6 B305D STAINING OF OTHER TISSUES (NORMAL KIDNEY,STOMACH AND PANCREAS) B305D IHC Tissue (Slide #) Reactivity CommentsStomach 1. Blk 85-568 cytomplasmic Grainy cytomplasmic staining (slide22) of glands below epithelium (not in neg control) 2. Blk 85-587cytomplasmic Graining staining of glands (slide 23) below epithelium,some background in negative control 3. Blk 85-1206 cytomplasmic Grainingstaining of glands (slide 24) below epithelium, lighter background innegative control 4. Blk 85-1225 cytomplasmic Marginal staining (slide25) 5. Blk 85-1426 cytomplasmic Grainy staining of glands (slide 26)below epithelium, some background in negative control Kidney 1. Blk00-7008 Inconclusive Staining of tubules; also (slide 27) (most likelypresent in neg control negative) (lighter) - mostly likely due toendogenous biotin 2. Blk 00-5638 Same as above Same as above (slide 28)3. Blk 00-1711 Same as above Same as above (slide 29) 4. Blk 00-3859Same as above Same as above (slide 30) 5. Blk 00-7651 Same as above Sameas above (slide 31) Pancreas 1. Blk Q965 Negative Non-specific stainingin (slide 32) negative control 2. Blk 00-2287 Negative Non-specificstaining in (slide 33) negative control 3. Blk 00-2790 NegativeNon-specific staining in (slide 34) negative control 4. Blk 00-6899Negative Non-specific staining in (slide 35) negative control 5. Blk00-7053 Negative Non-specific staining in (slide 36) negative control

[0415] In summary, B305D was only observed in less than 20% of breastcarcinomas. Staining was observed in half of the normal prostate sampleshowever, membrane staining was not detected in normal breast, in ovariancarcinomas or in normal pancreas, kidney, stomach or a panel of othernormal tissues.

Example 12 Analysis of Breast-Tumor Specific B305D Sequences

[0416] Numerous forms of the breast tumor antigen, B305D have beenisolated. To date, isoforms A (DNA SEQ ID NO: 291, 292, 296, 313, 314) Avariant (DNA SEQ ID NO: 299), B (DNA SEQ ID NO: 294, 297), and C (DNASEQ ID NO: 295, 301, 302, 303) have been identified. Using B305D genespecific 5′ and 3′ primers representing all known forms of B305D,specific forms of this gene expressed in breast tumors were amplified.Disclosed herein in SEQ ID NO: 341-348 are 4 D305D nucleotide sequencesand their corresponding amino acid sequences identified specifically inbreast tumors as described below.

[0417] Two PCR reactions were carried out using primers specific toB305D. The products were then analyzed and full-length sequences werecompiled. For the first reaction, primers were designed to regionscommon to all B305D forms near the 5′ and 3′ ends of the gene. Thesecond set of PCR reactions used primers specific to each of the startsites specific to each of the forms. Three 5′ primers were designed toamplify from the B305D A form, A form frameshift and C form start sites.3′ reverse primers were designed to a common region of all B305D forms,slightly upstream of the 3′ primer used in the first PCR reaction. PCRwas carried out using these primers and cDNA derived from breast tumorRNA numbers 443, 23B, and S76. All products were sequenced, analyzed andcompiled.

[0418] Two variants of the B305D A isoform were identified in the breasttumor samples. The nucleotide sequence of these 2 variants is set forthin SEQ ID NO: 341 and 342 and the corresponding amino acid sequence isset forth in SEQ ID NO: 345 and 346. One of these variants (SEQ ID NO:341) is identical to a previously identified variant of B305D A isoformdescribed in Example 1 and set forth in SEQ ID NO: 314. The othervariant (SEQ ID NO: 342) differs from SEQ ID NO: 314 by 2 base pairs andencodes an amino acid sequence (SEQ ID NO: 346) that differs by oneamino acid from the previously identified A isoform set forth in SEQ IDNO: 315.

[0419] Two new variants of the B305D C isoform were also identified fromthe breast tumor samples. The nucleotide sequence of these two variantsis provided in SEQ ID NO: 343 and 344 and the corresponding amino acidsequence is set forth in SEQ ID NO: 347 and 348. The 5′ end of the 2 Cisoform variants appears to be a truncated C isoform that is missing oneof the two 4 base pair repeats normally seen in the C isoform. The 3′end of these variants aligns well to the A isoforms. More specifically,there is a splice junction at around base 297. It is at this junctionwhere SEQ IDs 343 and 344 diverge from the standard C form and theremaining 3′ end being the A form. Upstream (5′ of) of this junction thesequence of B305D isoforms set forth in SEQ ID NO: 343 and 344 aremissing 111 base pairs of standard B305D C form respeat sequence. Thevariant set forth in SEQ ID 343 is the shortest, having an additional 6base pair deletion in the large missing repeat. Thus, in summary, SEQ IDNO: 343 and 344 begin with the ATG of the standard B305D C isoform. Thesequence continues as the C isoform for about 185 base pairs for SEQ IDNO: 344 and 179 base pairs for SEQ ID NO: 343. Both sequences then haveabout a 112 base pair deletion of repeat sequence just prior to thesplice junction. Following the splice junction, both variants follow theA form.

Example 13 Identification of CD4 T Cell Epitopes for B305D

[0420] This example demonstrates the identification of CD4+ T cellepitopes of the C form of B305D (full-length cDNA and amino acidsequence of B305D are set forth in SEQ ID NO: 301 and 304,respectively).

[0421] CD4+ T cell responses were generated using PBMC of normal donorsusing dendritic cells (DC) pulsed with overlapping 20-mer peptidesspanning the entire B305D C isoform protein. Briefly, CD4+ T cells werestimulated 3-4 times with DC pulsed with a mixture of overlappingpeptides in IMDM media containing IL-6 and IL-12 in the primarystimulation, and IL-2+IL-7 in all other stimulations. These lines weresubsequently assayed using a standard proliferation assay (measuringtritiated thymidine uptake) for reactivity with the priming peptides orrecombinant E. Coli derived B305D.

[0422] A number of different peptides elicited B305D specific T cells.These CD4+ T cell epitopes are contained in the following sequences:

[0423] VNKKDKQKRTALHLASANGNSEVVKLLLDR (SEQ ID NO: 349):

[0424] (peptides 34-46 corresponding to amino acids 166-195 of SEQ IDNO: 304).

[0425] ALHLASANGNSEVVKLLLDRRCQLNVLDNK (SEQ ID NO: 350)

[0426] (peptides 36-38 corresponding to amino acids 176-205 of SEQ IDNO: 304).

[0427] GSASIVSLLLEQNIDVSSQDLSGQT (SEQ ID NO: 351) (peptides 64-65corresponding to amino acids 316-340 of SEQ ID NO: 304).

[0428] CD4+ T cells recognizing these peptides also recognizerecombinant B305D protein, suggesting that these are naturally processedepitopes. Two of these lines (lines 31.9 and 31.10 recognizing peptidesset forth in SEQ ID NO: 349 and 350) also recognized mammalian sourcesof B305D including baculovirus protein, lysates from HEK cellstransiently transfected with B305D and lysates from cells infected withadenovirus expressing B305D.

[0429] Thus, these studies demonstrate that CD4+ T cell immunity toB305D can be elicited and identify the peptides set forth in SEQ ID NO:349-351 as immunogenic, naturally processed CD4+ T cell epitopes.

Example 14 Autoantibodies to B305D in Breast Cancer Sera and EpitopeMapping of the Antigenic Sites

[0430] Autoantibodies to specific B305D peptide epitopes were identifiedin the sera of breast cancer patients. Overlapping peptides spanning theentire B305D sequence (cDNA and amino acid sequence of the C form ofB305D set forth in SEQ ID NO: 301 and 304, respectively) weresynthesized and tested by ELISA with sera from patients with breastcancer to determine the presence of B305D-specific antibodies. Severalimmunoreactive regions were identified, including immunodominant regionsencompasssing the ankyrin repeat portion of the molecule.

[0431] Seventy-four 20-mer peptides overlapping by 15 amino acids,spanning the entire open reading frame of B305D were synthesized (aminoacid sequences set forth in SEQ ID NO: 352-425). These 74 peptides weretested in ELISA to evaluate which epitopes reacted with breast cancersera as well as control sera. Initially peptides were pooled and testedto locate regions of activity. Highest activity was obtained in peptides1-24 (SEQ ID NO: 352-375) and these were retested individually todetermine the specific epitopes. Peptides 3,5,6,11,13,19 and 20 (SEQ IDNO: 354, 356, 357, 362, 364, 370, 371, respectively) were then furthertested with a complete panel of 74 breast, 50 ovarian and 55 prostatecancer sera as well as controls. 18 of 74 breast cancer sera werereactive with one or more peptides. Both breast and ovarian cancer serashowed reactivity and active epitopes appeared located in the ankyrinrepeat regions of B305D. The amino acid sequence of the 3 ankyrin repeatsequences found in B305D are set forth in SEQ ID NO: 426-428 and arepresent within the overlapping peptides set forth in SEQ ID NO: 356-359,363-366, and 368-376, respectively.

[0432] Detection of autoantibodies to B305D in breast cancer seraindicates that such patients can elicit an immune response to specificepitopes and indicates that B305D can be used either alone or incombination with other breast tumor antigens as a target for vaccinedevelopment. Knowing that antibodies to B305D are present in the serumof breast cancer patients strengthens the potential use of this antigenas a vaccine target. In addition, detection of antibodies to B305D canbe used as a diagnostic for breast cancer alone or in combination withdetecting antibodies to other antigens, e.g., Her-2/neu or other tumorantigens. The presence of antibodies to B305D also indicates that B305Dantigen is present in serum and could be used as a target fordevelopment of a specific antigen detection assay.

Example 15 Aanalysis of cDNA Ecpression Using Microarray Technology

[0433] In additional studies, sequences disclosed herein are evaluatedfor overexpression in specific tumor tissues by microarray analysis.Using this approach, cDNA sequences are PCR amplified and their mRNAexpression profiles in tumor and normal tissues are examined using cDNAmicroarray technology essentially as described (Shena, M. et al., 1995Science 270:467-70). In brief, the clones are arrayed onto glass slidesas multiple replicas, with each location corresponding to a unique cDNAclone (as many as 5500 clones can be arrayed on a single slide, orchip). Each chip is hybridized with a pair of cDNA probes that arefluorescence-labeled with Cy3 and Cy5, respectively. Typically, 1 μg ofpolyA⁺ RNA is used to generate each cDNA probe. After hybridization, thechips are scanned and the fluorescence intensity recorded for both Cy3and Cy5 channels. There are multiple built-in quality control steps.First, the probe quality is monitored using a panel of ubiquitouslyexpressed genes. Secondly, the control plate also can include yeast DNAfragments of which complementary RNA may be spiked into the probesynthesis for measuring the quality of the probe and the sensitivity ofthe analysis. Currently, the technology offers a sensitivity of 1 in100,000 copies of mRNA. Finally, the reproducibility of this technologycan be ensured by including duplicated control cDNA elements atdifferent locations.

Example 16 Analysis of cDNA Expression Using Real-Time PCR

[0434] Real-time PCR (see Gibson et al., Genome Research 6:995-1001,1996; Heid et al., Genome Research 6:986-994,1996) is a technique thatevaluates the level of PCR product accumulation during amplification.This technique permits quantitative evaluation of mRNA levels inmultiple samples. Briefly, mRNA is extracted from tumor and normaltissue and cDNA is prepared using standard techniques. Real-time PCR isperformed, for example, using a Perkin Elmer/Applied Biosystems (FosterCity, Calif.) 7700 Prism instrument. Matching primers and fluorescentprobes are designed for genes of interest using, for example, the primerexpress program provided by Perkin Elmer/Applied Biosystems (FosterCity, Calif.). Optimal concentrations of primers and probes areinitially determined by those of ordinary skill in the art, and control(e.g., β-actin) primers and probes are obtained commercially from, forexample, Perkin Elmer/Applied Biosystems (Foster City, Calif.). Toquantitate the amount of specific RNA in a sample, a standard curve isgenerated using a plasmid containing the gene of interest. Standardcurves are generated using the Ct values determined in the real-timePCR, which are related to the initial cDNA concentration used in theassay. Standard dilutions ranging from 10-10⁶ copies of the gene ofinterest are generally sufficient. In addition, a standard curve isgenerated for the control sequence. This permits standardization ofinitial RNA content of a tissue sample to the amount of control forcomparison purposes.

[0435] An alternative real-time PCR procedure can be carried out asfollows: The first-strand cDNA to be used in the quantitative real-timePCR is synthesized from 20 μg of total RNA that is first treated withDNase I (e.g., Amplification Grade, Gibco BRL Life Technology,Gaitherburg, Md.), using Superscript Reverse Transcriptase (RT) (e.g.,Gibco BRL Life Technology, Gaitherburg, Md). Real-time PCR is performed,for example, with a GeneAmp™ 5700 sequence detection system (PEBiosystems, Foster City, Calif.). The 5700 system uses SYBR™ green, afluorescent dye that only intercalates into double stranded DNA, and aset of gene-specific forward and reverse primers. The increase influorescence is monitored during the whole amplification process. Theoptimal concentration of primers is determined using a checkerboardapproach and a pool of cDNAs from breast tumors is used in this process.The PCR reaction is performed in 25 μl volumes that include 2.5 μl ofSYBR green buffer, 2 μl of cDNA template and 2.5 μl each of the forwardand reverse primers for the gene of interest. The cDNAs used for RTreactions are diluted approximately 1:10 for each gene of interest and1:100 for the β-actin control. In order to quantitate the amount ofspecific cDNA (and hence initial mRNA) in the sample, a standard curveis generated for each run using the plasmid DNA containing the gene ofinterest. Standard curves are generated using the Ct values determinedin the real-time PCR which are related to the initial cDNA concentrationused in the assay. Standard dilution ranging from 20-2×10⁶ copies of thegene of interest are used for this purpose. In addition, a standardcurve is generated for β-actin ranging from 200 fg-2000 fg. This enablesstandardization of the initial RNA content of a tissue sample to theamount of β-actin for comparison purposes. The mean copy number for eachgroup of tissues tested is normalized to a constant amount of β-actin,allowing the evaluation of the over-expression levels seen with each ofthe genes.

Example 17 Peptide Priming of T-Helper Lines

[0436] Generation of CD4⁺ T helper lines and identification of peptideepitopes derived from tumor-specific antigens that ate capable of beingrecognized by CD4⁺ T cells in the context of HLA class II molecules, iscarried out as follows:

[0437] Fifteen-mer peptides overlapping by 10 amino acids, derived froma tumor-specific antigen, are generated using standard procedures.Dendritic cells (DC) are derived from PBMC of a normal donor usingGM-CSF and IL-4 by standard protocols. CD4⁺ T cells are generated fromthe same donor as the DC using MACS beads (Miltenyi Biotec, Auburn,Calif.) and negative selection. DC are pulsed overnight with pools ofthe 15-mer peptides, with each peptide at a final concentration of 0.25μg/ml. Pulsed DC are washed and plated at 1×10⁴ cells/well of 96-wellV-bottom plates and purified CD4⁺ T cells are added at 1×10⁵/well.Cultures are supplemented with 60 ng/ml IL-6 and 10 ng/ml IL-12 andincubated at 37° C. Cultures are restimulated as above on a weekly basisusing DC generated and pulsed as above as antigen presenting cells,supplemented with 5 ng/ml IL-7 and 10 U/mI IL-2. Following 4 in vitrostimulation cycles, resulting CD4⁺ T cell lines (each line correspondingto one well) are tested for specific proliferation and cytokineproduction in response to the stimulating pools of peptide with anirrelevant pool of peptides used as a control.

Example 18 Generation of Tumor-Specific CTL Lines Using in VitroWhole-Gene Priming

[0438] Using in vitro whole-gene priming with tumor antigen-vacciniainfected DC (see, for example, Yee et al, The Journal of Immunology,157(9):4079-86, 1996), human CTL lines are derived that specificallyrecognize autologous fibroblasts transduced with a specific tumorantigen, as determined by interferon-γ ELISPOT analysis. Specifically,dendritic cells (DC) are differentiated from monocyte cultures derivedfrom PBMC of normal human donors by growing for five days in RPMI mediumcontaining 10% human serum, 50 ng/ml human GM-CSF and 30 ng/ml humanIL-4. Following culture, DC are infected overnight with tumorantigen-recombinant vaccinia virus at a multiplicity of infection(M.O.I.) of five, and matured overnight by the addition of 3 μg/ml CD40ligand. Virus is then inactivated by UV irradiation. CD8+ T cells areisolated using a magnetic bead system, and priming cultures areinitiated using standard culture techniques. Cultures are restimulatedevery 7-10 days using autologous primary fibroblasts retrovirallytransduced with previously identified tumor antigens. Following fourstimulation cycles, CD8+ T cell lines are identified that specificallyproduce interferon-γ when stimulated with tumor antigen-transducedautologous fibroblasts. Using a panel of HLA-mismatched B-LCL linestransduced with a vector expressing a tumor antigen, and measuringinterferon-y production by the CTL lines in an ELISPOT assay, the HLArestriction of the CTL lines is determined.

Example 19 Generation and Characterization of Anti-Tumor AntigenMonoclonal Antibodies

[0439] Mouse monoclonal antibodies are raised against E. coli derivedtumor antigen proteins as follows: Mice are immunized with CompleteFreund's Adjuvant (CFA) containing 50 μg recombinant tumor protein,followed by a subsequent intraperitoneal boost with Incomplete Freund'sAdjuvant (IFA) containing 10 μg recombinant protein. Three days prior toremoval of the spleens, the mice are immunized intravenously withapproximately 50 μg of soluble recombinant protein. The spleen of amouse with a positive titer to the tumor antigen is removed, and asingle-cell suspension made and used for fusion to SP2/O myeloma cellsto generate B cell hybridomas. The supernatants from the hybrid clonesare tested by ELISA for specificity to recombinant tumor protein, andepitope mapped using peptides that spanned the entire tumor proteinsequence. The mAbs are also tested by flow cytometry for their abilityto detect tumor protein on the surface of cells stably transfected withthe cDNA encoding the tumor protein.

Example 20 Synthesis of Polypeptides

[0440] Polypeptides are synthesized on a Perkin Elmer/Applied BiosystemsDivision 430A peptide synthesizer using FMOC chemistry with HPTU(O-Benzotriazole-N,N,N′,N′-tetramethyluronium hexafluorophosphate)activation. A Gly-Cys-Gly sequence is attached to the amino terminus ofthe peptide to provide a method of conjugation, binding to animmobilized surface, or labeling of the peptide. Cleavage of thepeptides from the solid support is carried out using the followingcleavage mixture:trifluoroaceticacid:ethanedithiol:thioanisole:water:phenol (40:1:2:2:3).After cleaving for 2 hours, the peptides are precipitated in coldmethyl-t-butyl-ether. The peptide pellets are then dissolved in watercontaining 0.1% trifluoroacetic acid (TFA) and lyophilized prior topurification by C18 reverse phase HPLC. A gradient of 0%-60%acetonitrile (containing 0.1% TFA) in water (containing 0.1% TFA) isused to elute the peptides. Following lyophilization of the purefractions, the peptides are characterized using electrospray or othertypes of mass spectrometry and by amino acid analysis.

Example 21 Generation of B305D-Specific CTL Lines and Clones Using inVitro Whole-Gene Priming

[0441] This example describes the generation of B305D-specific CD8+ T 5lymphocytes from a normal donor and identification of the HLArestriction of two CD8+ T cell clones. B305D C isoform is a breast tumorantigen that is preferentially expressed in breast tumors as compared tonormal breast tissue. These experiments further confirm theimmunogenicity of the B305D protein and support its use as a target forvaccine and/or other immunotherapeutic approaches.

[0442] Standard in-vitro priming was established in 96-well platesgenerally as described in Example 18. More specifically, a total of 960cultures were established, using as APC DC infected with adenovirusexpressing B305D C isoform (SEQ ID NO: 301) for the initial stimulation,and autologous fibroblasts transduced to express the 5′ or 3′ ½ of B305DC isoform for 3 additional stimulations. T cell lines were screened byγ-IFN ELISPOT assays on fibroblasts expressing either the 5′ half (aminoacids 1-200 of SEQ ID NO: 304) or the 3′ half (amino acids 160-384 ofSEQ ID NO: 304) of B305D C isoform. Six T cell lines were identifiedthat recognized either the 5′ fragment (3B9, 7E5, and 8H8) or 3′fragment (4G2, 5E6, 7E10) of B305D C isoform. Clones were then generatedfrom lines 3B9, 5E6, and 8H8 and shown to recognize B305D by γ-IFNELISPOT assay. Antibody blocking γ-IFN ELISPOT assays were performed toidentify the relevant restricting alleles of each of the clones. Theactivity of 8H8 and 3B9 clones (3′ fragment specific) was specificallyblocked by pan class I and HLA-B/C blocking antibodies, and the activityof 5E6 clones was blocked by pan class I and HLA -A2 blockingantibodies. These results suggest that the restricting allele for the8H8 and 5E6 response is one of the B or C alleles of the donor, D385(B7, B35, Cw4, Cw7), and the restricting allele for the 3B9 clone is theHLA-A0205 allele expressed by D385. These results further suggest thatthere are at least 2 epitopes from B305D that are recognized by these Tcell clones.

[0443] In summary, these data demonstrate that precursor T cellsspecific for B305D C isoform exist that can be activated by vaccinationstrategies, and additionally indicate that naturally processed epitopesfrom B305D exist that can be used for both vaccination and immunemonitoring strategies.

[0444] From the foregoing it will be appreciated that, although specificembodiments of the invention have been described herein for purposes ofillustration, various modifications may be made without deviating fromthe spirit and scope of the invention. Accordingly, the invention is notlimited except as by the appended claims.

0 SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 428 <210> SEQ ID NO 1<211> LENGTH: 363 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 1 ttagagaccc aattgggacc taattgggac ccaaatttct caagtggagggagaactttt 60 gacgatttcc accggtatct cctcgtgggt attcagggag ctgcccagaaacctataaac 120 ttgtctaagg cgattgaagt cgtccagggg catgatgagt caccaggagtgtttttagag 180 cacctccagg aggcttatcg gatttacacc ccttttgacc tggcagcccccgaaaatagc 240 catgctctta atttggcatt tgtggctcag gcagccccag atagtaaaaggaaactccaa 300 aaactagagg gattttgctg gaatgaatac cagtcagctt ttagagatagcctaaaaggt 360 ttt 363 <210> SEQ ID NO 2 <211> LENGTH: 121 <212> TYPE:PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 2 Leu Glu Thr Gln LeuGly Pro Asn Trp Asp Pro Asn Phe Ser Ser Gly 1 5 10 15 Gly Arg Thr PheAsp Asp Phe His Arg Tyr Leu Leu Val Gly Ile Gln 20 25 30 Gly Ala Ala GlnLys Pro Ile Asn Leu Ser Lys Ala Ile Glu Val Val 35 40 45 Gln Gly His AspGlu Ser Pro Gly Val Phe Leu Glu His Leu Gln Glu 50 55 60 Ala Tyr Arg IleTyr Thr Pro Phe Asp Leu Ala Ala Pro Glu Asn Ser 65 70 75 80 His Ala LeuAsn Leu Ala Phe Val Ala Gln Ala Ala Pro Asp Ser Lys 85 90 95 Arg Lys LeuGln Lys Leu Glu Gly Phe Cys Trp Asn Glu Tyr Gln Ser 100 105 110 Ala PheArg Asp Ser Leu Lys Gly Phe 115 120 <210> SEQ ID NO 3 <211> LENGTH: 1080<212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 681, 685, 706, 720, 741, 752,758, 780, 789, 824, 840, 859,866, 884, 890, 905, 917, 926, 930, 951,957, 959, 962, 974, 980, 982, 988, 995, 996, 1007, 1010, 1025, 1040,1051, 1052, 1056, 1057, 1078 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 3 tcttagaatc ttcatacccc gaactcttgg gaaaacttta atcagtcacctacagtctac 60 cacccattta ggaggagcaa agctacctca gctcctccgg agccgttttaagatccccca 120 tcttcaaagc ctaacagatc aagcagctct ccggtgcaca acctgcgcccaggtaaatgc 180 caaaaaaggt cctaaaccca gcccaggcca ccgtctccaa gaaaactcaccaggagaaaa 240 gtgggaaatt gactttacag aagtaaaacc acaccgggct gggtacaaataccttctagt 300 actggtagac accttctctg gatggactga agcatttgct accaaaaacgaaactgtcaa 360 tatggtagtt aagtttttac tcaatgaaat catccctcga cgtgggctgcctgttgccat 420 agggtctgat aatggaacgg ccttcgcctt gtctatagtt taatcagtcagtaaggcgtt 480 aaacattcaa tggaagctcc attgtgccta tcgacccaga gctctgggcaagtagaacgc 540 atgaactgca ccctaaaaaa acactcttac aaaattaatc ttaaaaaccggtgttaattg 600 tgttagtctc cttcccttag ccctacttag agttaaggtg caccccttactgggctgggt 660 tctttacctt ttgaaatcat ntttnggaag gggctgccta tctttncttaactaaaaaan 720 gcccatttgg caaaaatttc ncaactaatt tntacgtncc tacgtctccccaacaggtan 780 aaaaatctnc tgcccttttc aaggaaccat cccatccatt cctnaacaaaaggcctgccn 840 ttcttccccc agttaactnt tttttnttaa aattcccaaa aaangaaccncctgctggaa 900 aaacnccccc ctccaanccc cggccnaagn ggaaggttcc cttgaatcccncccccncna 960 anggcccgga accnttaaan tngttccngg gggtnnggcc taaaagnccnatttggtaaa 1020 cctanaaatt ttttcttttn taaaaaccac nntttnnttt ttcttaaacaaaaccctntt 1080 <210> SEQ ID NO 4 <211> LENGTH: 1087 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature<222> LOCATION: 559, 574, 576, 581, 582, 587, 589, 593, 594, 609, 627,640,659, 668, 672, 677, 691, 713, 714, 732, 741, 812, 813, 823, 825,829, 838, 845, 849, 852, 855, 856, 859, 874, 876, 877, 892, 902, 907,916, 917, 938, 950, 951, 952, 953, 960 <223> OTHER INFORMATION: n =A,T,C or G <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:965, 974, 976, 978, 982, 996, 1005, 1012, 1049, 1058, 1073,1074, 1082,1084, 1086 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 4tctagagctg cgcctggatc ccgccacagt gaggagacct gaagaccaga gaaaacacag 60caagtaggcc ctttaaacta ctcacctgtg ttgtcttcta atttattctg ttttattttg 120tttccatcat tttaaggggt taaaatcatc ttgttcagac ctcagcatat aaaatgaccc 180atctgtagac ctcaggctcc aaccataccc caagagttgt ctggttttgt ttaaattact 240gccaggtttc agctgcagat atccctggaa ggaatattcc agattccctg agtagtttcc 300aggttaaaat cctataggct tcttctgttt tgaggaagag ttcctgtcag agaaaaacat 360gattttggat ttttaacttt aatgcttgtg aaacgctata aaaaaaattt tctaccccta 420gctttaaagt actgttagtg agaaattaaa attccttcag gaggattaaa ctgccatttc 480agttacccta attccaaatg ttttggtggt tagaatcttc tttaatgttc ttgaagaagt 540gttttatatt ttcccatcna gataaattct ctcncncctt nnttttntnt ctnntttttt 600aaaacggant cttgctccgt tgtccangct gggaattttn ttttggccaa tctccgctnc 660cttgcaanaa tnctgcntcc caaaattacc ncctttttcc cacctccacc ccnnggaatt 720acctggaatt anaggccccc nccccccccc cggctaattt gtttttgttt ttagtaaaaa 780acgggtttcc tgttttagtt aggatggccc anntctgacc ccntnatcnt ccccctcngc 840cctcnaatnt tnggnntang gcttaccccc cccngnngtt tttcctccat tnaaattttc 900tntggantct tgaatnncgg gttttccctt ttaaaccnat tttttttttn nnncccccan 960ttttncctcc cccntntnta angggggttt cccaanccgg gtccnccccc angtccccaa 1020tttttctccc cccccctctt ttttctttnc cccaaaantc ctatcttttc ctnnaaatat 1080cnantnt 1087 <210> SEQ ID NO 5 <211> LENGTH: 1010 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 311, 315, 318, 339, 341, 347, 361, 379, 391, 415, 417, 419,424, 430, 433, 454, 463, 465, 467, 476, 497, 499, 550, 562, 564, 587,591, 595, 597, 598, 612, 625, 631, 640, 641, 645, 648, 656, 661, 665,666, 670, 674, 675, 681, 682, 683 <223> OTHER INFORMATION: n = A,T,C orG <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 687, 688,692, 710, 721, 778, 788, 811, 820, 830, 860, 867,868, 871, 872, 889,892, 896, 897, 899, 904, 915, 936, 951, 960, 970, 986, 990, 1000 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 5 tctagaccaagaaatgggag gattttagag tgactgatga tttctctatc atctgcagtt 60 agtaaacattctccacagtt tatgcaaaaa gtaacaaaac cactgcagat gacaaacact 120 aggtaacacacatactatct cccaaatacc tacccacaag ctcaacaatt ttaaactgtt 180 aggatcactggctctaatca ccatgacatg aggtcaccac caaaccatca agcgctaaac 240 agacagaatgtttccactcc tgatccactg tgtgggaaga agcaccgaac ttacccactg 300 gggggcctgcntcanaanaa aagcccatgc ccccgggtnt ncctttnaac cggaacgaat 360 naacccaccatccccacanc tcctctgttc ntgggccctg catcttgtgg cctcntntnc 420 tttngggganacntggggaa ggtaccccat ttcnttgacc ccncnanaaa accccngtgg 480 ccctttgccctgattcncnt gggccttttc tcttttccct tttgggttgt ttaaattccc 540 aatgtccccngaaccctctc cntnctgccc aaaacctacc taaattnctc nctangnntt 600 ttcttggtgttncttttcaa aggtnacctt ncctgttcan ncccnacnaa aatttnttcc 660 ntatnntggncccnnaaaaa nnnatcnncc cnaattgccc gaattggttn ggtttttcct 720 nctgggggaaaccctttaaa tttccccctt ggccggcccc ccttttttcc cccctttnga 780 aggcaggnggttcttcccga acttccaatt ncaacagccn tgcccattgn tgaaaccctt 840 ttcctaaaattaaaaaatan ccggttnngg nnggcctctt tcccctccng gngggnngng 900 aaantccttaccccnaaaaa ggttgcttag cccccngtcc ccactccccc nggaaaaatn 960 aaccttttcnaaaaaaggaa tataantttn ccactccttn gttctcttcc 1010 <210> SEQ ID NO 6 <211>LENGTH: 950 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 199, 200, 209, 223, 224,236, 240, 241, 244, 248, 249, 262,263, 267, 268, 269, 270, 271, 272,273, 280, 281, 283, 285, 286, 287, 288, 289, 290, 291, 293, 295, 296,300, 302, 303, 309, 313, 314, 315, 316, 317, 318, 319, 320, 322, 323<223> OTHER INFORMATION: n = A,T,C or G <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 326, 327, 331, 332, 339, 342, 343, 344,346, 349, 352, 353,355, 356, 359, 360, 362, 363, 364, 367, 369, 371,375, 377, 378, 379, 383, 385, 387, 389, 390, 392, 396, 397, 399, 400,401, 402, 405, 406, 408, 409, 410, 412, 413, 414, 415 <223> OTHERINFORMATION: n = A,T,C or G <220> FEATURE: <221> NAME/KEY: misc_feature<222> LOCATION: 417, 419, 420, 423, 424, 428, 431, 433, 434, 435, 437,438, 439, 443, 447, 449, 450, 455, 456, 458, 459, 462, 465, 467, 469,472, 480, 481, 483, 484, 485, 486, 487, 488, 493, 494, 495, 496, 497,502, 505, 507, 508, 510, 512, 517, 518 <223> OTHER INFORMATION: n =A,T,C or G <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:520, 521, 524, 526, 531, 536, 538, 539, 543, 544, 548, 549, 550, 552,553, 555, 556, 557, 561, 563, 566, 570, 571, 572, 576, 577, 579, 580,582, 583, 585, 588, 590, 591, 592, 594, 597, 603, 606, 607, 614, 616,618, 620, 621, 622, 623 <223> OTHER INFORMATION: n = A,T,C or G <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 625, 628, 629,630, 632, 634, 637, 638, 641, 645, 651, 652, 653, 658, 659, 663, 664,668, 672, 673, 674, 678, 685, 689, 696, 700, 701, 702, 704, 705, 706,708, 710, 711, 712, 713, 715, 719, 722, 725, 727, 731, 734, 735, 737,739, 742 <223> OTHER INFORMATION: n = A,T,C or G <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 745, 748, 749, 751, 752, 754,755, 757, 759, 762, 765, 767, 769, 773, 774, 775, 778, 780, 783, 785,787, 790, 793, 797, 800, 803, 810, 812, 824, 828, 832, 836, 839, 843,844, 846, 848, 850, 852, 853, 855, 858, 859, 861, 864, 865, 866 <223>OTHER INFORMATION: n = A,T,C or G <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 868, 869, 872, 875, 880, 886, 889, 890,891, 892, 893, 895, 896, 901, 902, 906, 908, 913, 914, 916, 918, 921,924, 925, 930, 932, 935, 940 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 6 tctagagctc gcggccgcga gctctaatac gactcactat agggcgtcgactcgatctca 60 gctcactgca atctctgccc ccggggtcat gcgattctcc tgcctcagccttccaagtag 120 ctgggattac aggcgtgcaa caccacaccc ggctaatttt gtatttttaatagagatggg 180 gttttccctt gttggccann atggtctcna acccctgacc tcnngtgatccccccncccn 240 nganctcnna ctgctgggga tnnccgnnnn nnncctcccn ncncnnnnnnncncnntccn 300 tnntccttnc tcnnnnnnnn cnntcnntcc nncttctcnc cnnntnttntcnncnnccnn 360 cnnnccncnt ncccncnnnt tcncntncnn tntccnncnn nntcnncnnncnnnncntnn 420 ccnntacntc ntnnncnnnt ccntctntnn cctcnncnnt cnctncncnttntctcctcn 480 ntnnnnnnct ccnnnnntct cntcncnncn tncctcnntn nccncnccccncctcncnnc 540 ctnntttnnn cnncnnntcc ntnccnttcn nntccnntnn cnncntcncnnncnttnttc 600 ccnccnnttc cttncncntn nnntntcnnn cncntcnntc ntttnctcctnnntcccnnc 660 tcnnttcncc cnnntccncc ccccncctnt ctctcncccn nntnnntntnnnncntccnc 720 tntcncnttc ntcnntncnt tnctntcnnc nncnntncnc tnccntntntctnnntcncn 780 tcncntntcn ccntccnttn ctntctcctn tntccttccc ctcncctnctcnttcnccnc 840 ccnntntntn tnncnccnnt nctnnncnnc cntcntttcn tctctnctnnnnntnncctc 900 nncccntncc ctnntncnct nctnntaccn tnctnctccn tcttccttcc950 <210> SEQ ID NO 7 <211> LENGTH: 1086 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:501, 691, 711, 735, 751, 780, 810, 819, 826, 832, 849, 889, 890, 904,913, 920, 926, 937, 940, 953, 957, 960, 985, 993, 994, 1000, 1012, 1044,1060, 1063, 1080, 1081 <223> OTHER INFORMATION: n = A,T,C or G <400>SEQUENCE: 7 tctagagctc gcggccgcga gctcaattaa ccctcactaa agggagtcgactcgatcaga 60 ctgttactgt gtctatgtag aaagaagtag acataagaga ttccattttgttctgtacta 120 agaaaaattc ttctgccttg agatgctgtt aatctgtaac cctagccccaaccctgtgct 180 cacagagaca tgtgctgtgt tgactcaagg ttcaatggat ttagggctatgctttgttaa 240 aaaagtgctt gaagataata tgcttgttaa aagtcatcac cattctctaatctcaagtac 300 ccagggacac aatacactgc ggaaggccgc agggacctct gtctaggaaagccaggtatt 360 gtccaagatt tctccccatg tgatagcctg agatatggcc tcatgggaagggtaagacct 420 gactgtcccc cagcccgaca tcccccagcc cgacatcccc cagcccgacacccgaaaagg 480 gtctgtgctg aggaagatta ntaaaagagg aaggctcttt gcattgaagtaagaagaagg 540 ctctgtctcc tgctcgtccc tgggcaataa aatgtcttgg tgttaaacccgaatgtatgt 600 tctacttact gagaatagga gaaaacatcc ttagggctgg aggtgagacaccctggcggc 660 atactgctct ttaatgcacg agatgtttgt ntaattgcca tccagggccancccctttcc 720 ttaacttttt atganacaaa aactttgttc ncttttcctg cgaacctctccccctattan 780 cctattggcc tgcccatccc ctccccaaan ggtgaaaana tgttcntaaatncgagggaa 840 tccaaaacnt tttcccgttg gtcccctttc caaccccgtc cctgggccnntttcctcccc 900 aacntgtccc ggntccttcn ttcccncccc cttcccngan aaaaaaccccgtntganggn 960 gccccctcaa attataacct ttccnaaaca aannggttcn aaggtggtttgnttccggtg 1020 cggctggcct tgaggtcccc cctncacccc aatttggaan ccngttttttttattgcccn 1080 ntcccc 1086 <210> SEQ ID NO 8 <211> LENGTH: 1177 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 1, 4, 20, 21, 31, 278, 314, 332, 359, 371,373, 375, 376, 524, 537, 556, 557, 579, 583, 590, 591, 598, 623, 625,648, 700, 703, 719, 738, 742, 746, 749, 751, 752, 800, 808, 820, 821,824, 835, 838, 845, 851, 856, 864, 865, 879, 888 <223> OTHERINFORMATION: n = A,T,C or G <220> FEATURE: <221> NAME/KEY: misc_feature<222> LOCATION: 911, 920, 926, 935, 945, 950, 952, 956, 969, 972, 977,981, 992, 999, 1023, 1024, 1032, 1038, 1039, 1040, 1062, 1069, 1075,1084, 1089, 1104, 1119, 1123, 1131, 1143, 1146, 1152, 1165, 1169, 1172,1176 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 8nccntttaga tgttgacaan ntaaacaagc ngctcaggca gctgaaaaaa gccactgata 60aagcatcctg gagtatcaga gtttactgtt agatcagcct catttgactt cccctcccac 120atggtgttta aatccagcta cactacttcc tgactcaaac tccactattc ctgttcatga 180ctgtcaggaa ctgttggaaa ctactgaaac tggccgacct gatcttcaaa atgtgcccct 240aggaaaggtg gatgccaccg tgttcacaga cagtaccncc ttcctcgaga agggactacg 300aggggccggt gcanctgtta ccaaggagac tnatgtgttg tgggctcagg ctttaccanc 360aaacacctca ncncnnaagg ctgaattgat cgccctcact caggctctcg gatggggtaa 420gggatattaa cgttaacact gacagcaggt acgcctttgc tactgtgcat gtacgtggag 480ccatctacca ggagcgtggg ctactcactc ggcaggtggc tgtnatccac tgtaaangga 540catcaaaagg aaaacnnggc tgttgcccgt ggtaaccana aanctgatcn ncagctcnaa 600gatgctgtgt tgactttcac tcncncctct taaacttgct gcccacantc tcctttccca 660accagatctg cctgacaatc cccatactca aaaaaaaaan aanactggcc ccgaacccna 720accaataaaa acggggangg tnggtnganc nncctgaccc aaaaataatg gatcccccgg 780gctgcaggaa ttcaattcan ccttatcnat acccccaacn nggngggggg ggccngtncc 840cattncccct ntattnattc tttnnccccc cccccggcnt cctttttnaa ctcgtgaaag 900ggaaaacctg ncttaccaan ttatcncctg gaccntcccc ttccncggtn gnttanaaaa 960aaaagcccnc antcccntcc naaatttgca cngaaaggna aggaatttaa cctttatttt 1020ttnntccttt antttgtnnn ccccctttta cccaggcgaa cngccatcnt ttaanaaaaa 1080aaanagaang tttatttttc cttngaacca tcccaatana aancacccgc nggggaacgg 1140ggnggnaggc cnctcacccc ctttntgtng gngggnc 1177 <210> SEQ ID NO 9 <211>LENGTH: 1146 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 1, 4, 5, 8, 9, 348, 706,742, 745, 751, 758, 772, 793, 819, 842, 846, 860, 866, 886, 889, 911,939, 945, 955, 960, 982, 999, 1002, 1005, 1009, 1010, 1033, 1047, 1049,1055, 1058, 1069, 1074, 1079, 1081, 1104, 1105, 1111, 1116, 1118 <223>OTHER INFORMATION: n = A,T,C or G <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 1121, 1130, 1135, 1136, 1146 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 9 nccnnttnnt gatgttgtctttttggcctc tctttggata ctttccctct cttcagaggt 60 gaaaagggtc aaaaggagctgttgacagtc atcccaggtg ggccaatgtg tccagagtac 120 agactccatc agtgaggtcaaagcctgggg cttttcagag aagggaggat tatgggtttt 180 ccaattatac aagtcagaagtagaaagaag ggacataaac caggaagggg gtggagcact 240 catcacccag agggacttgtgcctctctca gtggtagtag aggggctact tcctcccacc 300 acggttgcaa ccaagaggcaatgggtgatg agcctacagg ggacatancc gaggagacat 360 gggatgaccc taagggagtaggctggtttt aaggcggtgg gactgggtga gggaaactct 420 cctcttcttc agagagaagcagtacagggc gagctgaacc ggctgaaggt cgaggcgaaa 480 acacggtctg gctcaggaagaccttggaag taaaattatg aatggtgcat gaatggagcc 540 atggaagggg tgctcctgaccaaactcagc cattgatcaa tgttagggaa actgatcagg 600 gaagccggga atttcattaacaacccgcca cacagcttga acattgtgag gttcagtgac 660 ccttcaaggg gccactccactccaactttg gccattctac tttgcnaaat ttccaaaact 720 tcctttttta aggccgaatccntantccct naaaaacnaa aaaaaatctg cncctattct 780 ggaaaaggcc cancccttaccaggctggaa gaaattttnc cttttttttt tttttgaagg 840 cntttnttaa attgaacctnaattcncccc cccaaaaaaa aacccnccng gggggcggat 900 ttccaaaaac naattcccttaccaaaaaac aaaaacccnc ccttnttccc ttccnccctn 960 ttcttttaat tagggagagatnaagccccc caatttccng gnctngatnn gtttcccccc 1020 cccccatttt ccnaaactttttcccancna ggaanccncc ctttttttng gtcngattna 1080 ncaaccttcc aaaccatttttccnnaaaaa ntttgntngg ngggaaaaan acctnntttt 1140 atagan 1146 <210> SEQID NO 10 <211> LENGTH: 545 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 10 cttcattggg tacgggcccc ctcgaggtcg acggtatcgataagcttgat atcgaattcc 60 tgcagcccgg gggatccact agttctagag tcaggaagaaccaccaacct tcctgatttt 120 tattggctct gagttctgag gccagttttc ttcttctgttgagtatgcgg gattgtcagg 180 cagatctggc tgtggaaagg agactgtggg cagcaagtttagaggcgtga ctgaaagtca 240 cactgcatct tgagctgctg aatcagcttt ctggttaccacgggcaacag ccgtgttttc 300 cttttgatgt cctttacagt ggattacagc cacctgctgaggtgagtagc ccacgctcct 360 ggtagatggc tccacgtaca tgcacagtag caaaggcgtacctgctgtca gtgttaacgt 420 taatatcctt accccatcgg agagcctgag tgagggcgatcaattcagcc cttttgtgct 480 gaggtgtttg ctggttaagc cctgaaccca caacacatctgtctccatgg taacagctgc 540 accgg 545 <210> SEQ ID NO 11 <211> LENGTH: 196<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 11tctcctaggc tgggcacagt ggctcatacc tgtaatcctg accgtttcag aggctcaggt 60ggggggatcg cttgagccca agatttcaag actagtctgg gtaacatagt gagaccctat 120ctctacgaaa aaataaaaaa atgagcctgg tgtagtggca cacaccagct gaggagggag 180aatcgagcct aggaga 196 <210> SEQ ID NO 12 <211> LENGTH: 388 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 82, 162, 287 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 12 tctcctaggc ttgggggctc tgactagaaattcaaggaac ctgggattca agtccaactg 60 tgacaccaac ttacactgtg gnctccaataaactgcttct ttcctattcc ctctctatta 120 aataaaataa ggaaaacgat gtctgtgtatagccaagtca gntatcctaa aaggagatac 180 taagtgacat taaatatcag aatgtaaaacctgggaacca ggttcccagc ctgggattaa 240 actgacagca agaagactga acagtactactgtgaaaagc ccgaagnggc aatatgttca 300 ctctaccgtt gaaggatggc tgggagaatgaatgctctgt cccccagtcc caagctcact 360 tactatacct cctttatagc ctaggaga 388<210> SEQ ID NO 13 <211> LENGTH: 337 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 13 tagtagttgc ctataatcat gtttctcattattttcacat tttattaacc aatttctgtt 60 taccctgaaa aatatgaggg aaatatatgaaacagggagg caatgttcag ataattgatc 120 acaagatatg atttctacat cagatgctctttcctttcct gtttatttcc tttttatttc 180 ggttgtgggg tcgaatgtaa tagctttgtttcaagagaga gttttggcag tttctgtagc 240 ttctgacact gctcatgtct ccaggcatctatttgcactt taggaggtgt cgtgggagac 300 tgagaggtct attttttcca tatttgggcaactacta 337 <210> SEQ ID NO 14 <211> LENGTH: 571 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 435, 441, 451, 456, 462, 479, 488, 489, 509, 568 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 14 tagtagttgc catacagtgcctttccattt atttaacccc cacctgaacg gcataaactg 60 agtgttcagc tggtgttttttactgtaaac aataaggaga ctttgctctt catttaaacc 120 aaaatcatat ttcatattttacgctcgagg gtttttaccg gttccttttt acactcctta 180 aaacagtttt taagtcgtttggaacaagat attttttctt tcctggcagc ttttaacatt 240 atagcaaatt tgtgtctgggggactgctgg tcactgtttc tcacagttgc aaatcaaggc 300 atttgcaacc aagaaaaaaaaatttttttg ttttatttga aactggaccg gataaacggt 360 gtttggagcg gctgctgtatatagttttaa atggtttatt gcacctcctt aagttgcact 420 tatgtggggg ggggnttttgnatagaaagt ntttantcac anagtcacag ggacttttnt 480 cttttggnna ctgagctaaaaagggctgnt tttcgggtgg gggcagatga aggctcacag 540 gaggcctttc tcttagaggggggaactnct a 571 <210> SEQ ID NO 15 <211> LENGTH: 548 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature<222> LOCATION: 224, 291, 326, 376, 388, 394, 428, 433, 507, 514 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 15 tatatatttaataacttaaa tatattttga tcacccactg gggtgataag acaatagata 60 taaaagtatttccaaaaagc ataaaaccaa agtatcatac caaaccaaat tcatactgct 120 tcccccacccgcactgaaac ttcaccttct aactgtctac ctaaccaaat tctacccttc 180 aagtctttggtgcgtgctca ctactctttt tttttttttt tttnttttgg agatggagtc 240 tggctgtgcagcccaggggt ggagtacaat ggcacaacct cagctcactg naacctccgc 300 ctcccaggttcatgagattc tcctgnttca gccttcccag tagctgggac tacaggtgtg 360 catcaccatgcctggntaat cttttttngt tttngggtag agatgggggt tttacatgtt 420 ggccaggntggtntcgaact cctgacctca agtgatccac ccacctcagg ctcccaaagt 480 gctaggattacagacatgag ccactgngcc cagncctggt gcatgctcac ttctctaggc 540 aactacta 548<210> SEQ ID NO 16 <211> LENGTH: 638 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:471, 488 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 16ttccgttatg cacatgcaga atattctatc ggtacttcag ctattactca ttttgatggc 60gcaatccgag cctatcctca agatgagtat ttagaaagaa ttgatttagc gatagaccaa 120gctggtaagc actctgacta cacgaaattg ttcagatgtg atggatttat gacagttgat 180ctttggaaga gattattaag tgattatttt aaagggaatc cattaattcc agaatatctt 240ggtttagctc aagatgatat agaaatagaa cagaaagaga ctacaaatga agatgtatca 300ccaactgata ttgaagagcc tatagtagaa aatgaattag ctgcatttat tagccttaca 360catagcgatt ttcctgatga atcttatatt cagccatcga catagcatta cctgatgggc 420aaccttacga ataatagaaa ctgggtgcgg ggctattgat gaattcatcc ncagtaaatt 480tggatatnac aaaatataac tcgattgcat ttggatgatg gaatactaaa tctggcaaaa 540gtaactttgg agctactagt aacctctctt tttgagatgc aaaattttct tttagggttt 600cttattctct actttacgga tattggagca taacggga 638 <210> SEQ ID NO 17 <211>LENGTH: 286 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:17 actgatggat gtcgccggag gcgaggggcc ttatctgatg ctcggctgcc tgttcgtgat 60gtgcgcggcg attgggctgt ttatctcaaa caccgccacg gcggtgctga tggcgcctat 120tgccttagcg gcggcgaagt caatgggcgt ctcaccctat ccttttgcca tggtggtggc 180gatggcggct tcggcggcgt ttatgacccc ggtctcctcg ccggttaaca ccctggtgct 240tggccctggc aagtactcat ttagcgattt tgtcaaaata ggcgtg 286 <210> SEQ ID NO18 <211> LENGTH: 262 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 184, 234, 240<223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 18 tcggtcatagcagccccttc ttctcaattt catctgtcac taccctggtg tagtatctca 60 tagccttacatttttatagc ctcctccctg gtctgtcttt tgattttcct gcctgtaatc 120 catatcacacataactgcaa gtaaacattt ctaaagtgtg gttatgctca tgtcactcct 180 gtgncaagaaatagtttcca ttaccgtctt aataaaattc ggatttgttc tttnctattn 240 tcactcttcacctatgaccg aa 262 <210> SEQ ID NO 19 <211> LENGTH: 261 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 19 tcggtcatag caaagccagtggtttgagct ctctactgtg taaactccta aaccaaggcc 60 atttatgata aatggtggcaggatttttat tataaacatg tacccatgca aatttcctat 120 aactctgaga tatattcttctacatttaaa caataaaaat aatctatttt taaaagccta 180 atttgcgtag ttaggtaagagtgtttaatg agagggtata aggtataaat caccagtcaa 240 cgtttctctg cctatgaccg a261 <210> SEQ ID NO 20 <211> LENGTH: 294 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:194, 274, 283, 294 <223> OTHER INFORMATION: n = A,T,C or G <400>SEQUENCE: 20 tacaacgagg cgacgtcggt aaaatcggac atgaagccac cgctggtcttttcgtccgag 60 cgataggcgc cggccagcca gcggaacggt tgcccggatg gcgaagcgagccggagttct 120 tcggactgag tatgaatctt gttgtgaaaa tactcgccgc cttcgttcgacgacgtcgcg 180 tcgaaatctt cganctcctt acgatcgaag tcttcgtggg cgacgatcgcggtcagttcc 240 gccccaccga aatcatggtt gagccggatg ctgnccccga agncctcgtttgtn 294 <210> SEQ ID NO 21 <211> LENGTH: 208 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 116, 132, 140, 160, 164, 191, 197, 199 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 21 ttggtaaagg gcatggacgcagacgcctga cgtttggctg aaaatctttc attgattcgt 60 atcaatgaat aggaaaattcccaaagaggg aatgtcctgt tgctcgccag tttttntgtt 120 gttctcatgg anaaggcaangagctcttca gactattggn attntcgttc ggtcttctgc 180 caactagtcg ncttgcnangatcttcat 208 <210> SEQ ID NO 22 <211> LENGTH: 287 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 1, 4, 25, 121, 168, 207, 212 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 22 nccnttgagc tgagtgattg agatntgtaatggttgtaag ggtgattcag gcggattagg 60 gtggcgggtc acccggcagt gggtctcccgacaggccagc aggatttggg gcaggtacgg 120 ngtgcgcatc gctcgactat atgctatggcaggcgagccg tggaaggngg atcaggtcac 180 ggcgctggag ctttccacgg tccatgnattgngatggctg ttctaggcgg ctgttgccaa 240 gcgtgatggt acgctggctg gagcattgatttctggtgcc aaggtgg 287 <210> SEQ ID NO 23 <211> LENGTH: 204 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 40, 121, 131, 162, 184, 197 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 23 ttgggtaaag ggagcaaggagaaggcatgg agaggctcan gctggtcctg gcctacgact 60 gggccaagct gtcgccggggatggtggaga actgaagcgg gacctcctcg aggtcctccg 120 ncgttacttc nccgtccaggaggagggtct ttccgtggtc tnggaggagc ggggggagaa 180 gatnctcctc atggtcnacatccc 204 <210> SEQ ID NO 24 <211> LENGTH: 264 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 171, 206 <223> OTHER INFORMATION: n = A,T,C or G <400>SEQUENCE: 24 tggattggtc aggagcgggt agagtggcac cattgagggg atattcaaaaatattatttt 60 gtcctaaatg atagttgctg agtttttctt tgacccatga gttatattggagtttatttt 120 ttaactttcc aatcgcatgg acatgttaga cttattttct gttaatgattnctattttta 180 ttaaattgga tttgagaaat tggttnttat tatatcaatt tttggtatttgttgagtttg 240 acattatagc ttagtatgtg acca 264 <210> SEQ ID NO 25 <211>LENGTH: 376 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 103, 111, 192, 196, 199,220, 224, 230, 251, 268, 283, 317, 352, 370, 374 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 25 ttacaacgag gggaaactccgtctctacaa aaattaaaaa attagccagg tgtggtggtg 60 tgcacccgca atcccagctacttgggaggt tgagacacaa gantcaccta natgtgggag 120 gtcaaggttg catgagtcatgattgtgcca ctgcactcca gcctgggtga cagaccgaga 180 ccctgcctca anaganaangaataggaagt tcagaaatcn tggntgtggn gcccagcaat 240 ctgcatctat ncaacccctgcaggcaangc tgatgcagcc tangttcaag agctgctgtt 300 tctggaggca gcagttngggcttccatcca gtatcacggc cacactcgca cnagccatct 360 gtcctccgtn tgtnac 376<210> SEQ ID NO 26 <211> LENGTH: 372 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:231, 312, 340 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 26ttacaacgag gggaaactcc gtctctacaa aaattaaaaa attagccagg tgtggtggtg 60tgcacctgta atcccagcta cttgggcggc tgagacacaa gaaccaccta aatgtgggag 120ggtcaaggtt gcatgagtca tgatcgcgcc actgcactcc agcctgggtg acagactgag 180accctgcctc aaaagaaaaa gaataggaag ttcagaaacc ctgggtgtgg ngcccagcaa 240tctgcattta aacaatccct gcaggcaatg ctgatgcagc ctaagttcaa gagctgctgt 300tctggaggca gnagtaaggg cttccatcca gcatcacggn caacactgca aaagcacctg 360tcctcgttgg ta 372 <210> SEQ ID NO 27 <211> LENGTH: 477 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 27 ttctgtccac atctacaagttttatttatt ttgtgggttt tcagggtgac taagtttttc 60 cctacattga aaagagaagttgctaaaagg tgcacaggaa atcatttttt taagtgaata 120 tgataatatg ggtccgtgcttaatacaact gagacatatt tgttctctgt ttttttagag 180 tcacctctta aagtccaatcccacaatggt gaaaaaaaaa tagaaagtat ttgttctacc 240 tttaaggaga ctgcagggattctccttgaa aacggagtat ggaatcaatc ttaaataaat 300 atgaaattgg ttggtcttctgggataagaa attcccaact cagtgtgctg aaattcacct 360 gacttttttt gggaaaaaatagtcgaaaat gtcaatttgg tccataaaat acatgttact 420 attaaaagat atttaaagacaaattctttc agagctctaa gattggtgtg gacagaa 477 <210> SEQ ID NO 28 <211>LENGTH: 438 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 4, 16, 30, 255, 413 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 28 tctncaacctcttgantgtc aaaaaccttn taggctatct ctaaaagctg actggtattc 60 attccagcaaaatccctcta gtttttggag tttcctttta ctatctgggg ctgcctgagc 120 cacaaatgccaaattaagag catggctatt ttcgggggct gacaggtcaa aaggggtgta 180 aatccgataagcctcctgga ggtgctctaa aaacactcct ggtgactcat catgcccctg 240 gacgacttcaatcgncttag acaagtttat aggtttctgg gcagctccct gaatacccac 300 gaggagataccggtggaaat cgtcaaaagt tctccctcca cttgagaaat ttgggtccca 360 attaggtcccaattgggtct ctaatcacta ttcctctagc ttcctcctcc ggnctattgg 420 ttgatgtgaggttgaaga 438 <210> SEQ ID NO 29 <211> LENGTH: 620 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 391, 481, 483, 490, 497, 510, 527, 532, 540, 545, 593, 612<223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 29 aagagggtaccagccccaag ccttgacaac ttccataggg tgtcaagcct gtgggtgcac 60 agaagtcaaaaattgagttt tgggatcctc agcctagatt tcagaggata taaagaaaca 120 cctaacacctagatattcag acaaaagttt actacaggga tgaagctttc acggaaaacc 180 tctactaggaaagtacagaa gagaaatgtg ggtttggagc ccccaaacag aatcccctct 240 agaacactgcctaatgaaac tgtgagaaga tggccactgt catccagaca ccagaatgat 300 agacccaccaaaaacttatg ccatattgcc tataaaacct acagacactc aatgccagcc 360 ccatgaaaaaaaaactgaga agaagactgt nccctacaat gccaccggag cagaactgcc 420 ccaggccatggaagcacagc tcttatatca atgtgacctg gatgttgaga catggaatcc 480 nangaaatcnttttaanact tccacggttn aatgactgcc ctattanatt cngaacttan 540 atccnggcctgtgacctctt tgctttggcc attccccctt tttggaatgg ctnttttttt 600 cccatgcctgtnccctctta 620 <210> SEQ ID NO 30 <211> LENGTH: 100 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 30 ttacaacgag ggggtcaatgtcataaatgt cacaataaaa caatctcttc tttttttttt 60 tttttttttt tttttttttttttttttttt tttttttttt 100 <210> SEQ ID NO 31 <211> LENGTH: 762 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 626, 652, 662, 715, 736 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 31 tagtctatgc gccggacagagcagaattaa attggaagtt gccctccgga ctttctaccc 60 acactcttcc tgaaaagagaaagaaaagag gcaggaaaga ggttaggatt tcattttcaa 120 gagtcagcta attaggagagcagagtttag acagcagtag gcaccccatg atacaaacca 180 tggacaaagt ccctgtttagtaactgccag acatgatcct gctcaggttt tgaaatctct 240 ctgcccataa aagatggagagcaggagtgc catccacatc aacacgtgtc caagaaagag 300 tctcagggag acaagggtatcaaaaaacaa gattcttaat gggaaggaaa tcaaaccaaa 360 aaattagatt tttctctacatatatataat atacagatat ttaacacatt attccagagg 420 tggctccagt ccttggggcttgagagatgg tgaaaacttt tgttccacat taacttctgc 480 tctcaaattc tgaagtatatcagaatggga caggcaatgt tttgctccac actggggcac 540 agacccaaat ggttctgtgcccgaagaaga gaagcccgaa agacatgaag gatgcttaag 600 gggggttggg aaagccaaattggtantatc ttttcctcct gcctgtgttc cngaagtctc 660 cnctgaagga attcttaaaaccctttgtga ggaaatgccc ccttaccatg acaantggtc 720 ccattgcttt tagggngatggaaacaccaa gggttttgat cc 762 <210> SEQ ID NO 32 <211> LENGTH: 276 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 32 tagtctatgcgtgtattaac ctcccctccc tcagtaacaa ccaaagaggc aggagctgtt 60 attaccaaccccattttaca gatgcatcaa taatgacaga gaagtgaagt gacttgcgca 120 cacaaccagtaaattggcag agtcagattt gaatccatgg agtctggtct gcactttcaa 180 tcaccgaataccctttctaa gaaacgtgtg ctgaatgagt gcatggataa atcagtgtct 240 actcaacatctttgcctaga tatcccgcat agacta 276 <210> SEQ ID NO 33 <211> LENGTH: 477<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 33tagtagttgc caaatatttg aaaatttacc cagaagtgat tgaaaacttt ttggaaacaa 60aaacaaataa agccaaaagg taaaataaaa atatctttgc actctcgtta ttacctatcc 120ataacttttt caccgtaagc tctcctgctt gttagtgtag tgtggttata ttaaactttt 180tagttattat tttttattca cttttccact agaaagtcat tattgattta gcacacatgt 240tgatctcatt tcattttttc tttttatagg caaaatttga tgctatgcaa caaaaatact 300caagcccatt atcttttttc cccccgaaat ctgaaaattg caggggacag agggaagtta 360tcccattaaa aaattgtaaa tatgttcagt ttatgtttaa aaatgcacaa aacataagaa 420aattgtgttt acttgagctg ctgattgtaa gcagttttat ctcaggggca actacta 477 <210>SEQ ID NO 34 <211> LENGTH: 631 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 34 tagtagttgc caattcagat gatcagaaat gctgctttcctcagcattgt cttgttaaac 60 cgcatgccat ttggaacttt ggcagtgaga agccaaaaggaagaggtgaa tgacatatat 120 atatatatat attcaatgaa agtaaaatgt atatgctcatatactttcta gttatcagaa 180 tgagttaagc tttatgccat tgggctgctg catattttaatcagaagata aaagaaaatc 240 tgggcatttt tagaatgtga tacatgtttt tttaaaactgttaaatatta tttcgatatt 300 tgtctaagaa ccggaatgtt cttaaaattt actaaaacagtattgtttga ggaagagaaa 360 actgtactgt ttgccattat tacagtcgta caagtgcatgtcaagtcacc cactctctca 420 ggcatcagta tccacctcat agctttacac attttgacggggaatattgc agcatcctca 480 ggcctgacat ctgggaaagg ctcagatcca cctactgctccttgctcgtt gatttgtttt 540 aaaatattgt gcctggtgtc acttttaagc cacagccctgcctaaaagcc agcagagaac 600 agaacccgca ccattctata ggcaactact a 631 <210>SEQ ID NO 35 <211> LENGTH: 578 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 35 tagtagttgc catcccatat tacagaaggc tctgtatacatgacttattt ggaagtgatc 60 tgttttctct ccaaacccat ttatcgtaat ttcaccagtcttggatcaat cttggtttcc 120 actgatacca tgaaacctac ttggagcaga cattgcacagttttctgtgg taaaaactaa 180 aggtttattt gctaagctgt catcttatgc ttagtattttttttttacag tggggaattg 240 ctgagattac attttgttat tcattagata ctttgggataacttgacact gtcttctttt 300 tttcgctttt aattgctatc atcatgcttt tgaaacaagaacacattagt cctcaagtat 360 tacataagct tgcttgttac gcctggtggt ttaaaggactatctttggcc tcaggttcac 420 aagaatgggc aaagtgtttc cttatgttct gtagttctcaataaaagatt gccaggggcc 480 gggtactgtg gctcgcactg taatcccagc actttgggaagctgaggctg gcggatcatg 540 ttagggcagg tgttcgaaac cagcctgggc aactacta 578<210> SEQ ID NO 36 <211> LENGTH: 583 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 36 tagtagttgc ctgtaatccc agcaactcaggaggctgggg caggagaatc agttgaacct 60 gggaggcaga agttgtaatt agcaaagatcgcaccattgc acttcagcct gggcaacaag 120 agtgagattc catctcaaaa acaaaaaaaagaaaaagaaa agaaaaggaa aaaacgtata 180 aacccagcca aaacaaaatg atcattcttttaataagcaa gactaattta atgtgtttat 240 ttaatcaaag cagttgaatc ttctgagttattggtgaaaa tacccatgta gttaatttag 300 ggttcttact tgggtgaacg tttgatgttcacaggttata aaatggttaa caaggaaaat 360 gatgcataaa gaatcttata aactactaaaaataaataaa atataaatgg ataggtgcta 420 tggatggagt ttttgtgtaa tttaaaatcttgaagtcatt ttggatgctc attggttgtc 480 tggtaatttc cattaggaaa aggttatgatatggggaaac tgtttctgga aattgcggaa 540 tgtttctcat ctgtaaaatg ctagtatctcagggcaacta cta 583 <210> SEQ ID NO 37 <211> LENGTH: 716 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature<222> LOCATION: 15, 669, 673, 678, 686, 704 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 37 gatctactag tcatntggat tctatccatggcagctaagc ctttctgaat ggattctact 60 gctttcttgt tctttaatcc agacccttatatatgtttat gttcacaggc agggcaatgt 120 ttagtgaaaa caattctaaa ttttttattttgcattttca tgctaatttc cgtcacactc 180 cagcaggctt cctgggagaa taaggagaaatacagctaaa gacattgtcc ctgcttactt 240 acagcctaat ggtatgcaaa accacttcaataaagtaaca ggaaaagtac taaccaggta 300 gaatggacca aaactgatat agaaaaatcagaggaagaga ggaacaaata tttactgagt 360 cctagaatgt acaaggcttt ttaattacatattttatgta aggcctgcaa aaaacaggtg 420 agtaatcaac atttgtccca ttttacatataaggaaactg aagcttaaat tgaataattt 480 aatgcataga ttttatagtt agaccatgttcaggtcccta tgttatactt actagctgta 540 tgaatatgag aaaataattt tgttattttcttggcatcag tattttcatc tgcaaaataa 600 agctaaagtt atttagcaaa cagtcagcatagtgcctgat acatagtagg tgctccaaac 660 atgattacnc tantattngg tattanaaaaatccaatata ggcntggata aaaccg 716 <210> SEQ ID NO 38 <211> LENGTH: 688<212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 260 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 38 ttctgtccac atatcatccc actttaattgttaatcagca aaactttcaa tgaaaaatca 60 tccattttaa ccaggatcac accaggaaactgaaggtgta ttttttttta ccttaaaaaa 120 aaaaaaaaaa accaaacaaa ccaaaacagattaacagcaa agagttctaa aaaatttaca 180 tttctcttac aactgtcatt cagagaacaatagttcttaa gtctgttaaa tcttggcatt 240 aacagagaaa cttgatgaan agttgtacttggaatattgt ggattttttt ttttgtctaa 300 tctcccccta ttgttttgcc aacagtaatttaagtttgtg tggaacatcc ccgtagttga 360 agtgtaaaca atgtatagga aggaatatatgataagatga tgcatcacat atgcattaca 420 tgtagggacc ttcacaactt catgcactcagaaaacatgc ttgaagagga ggagaggacg 480 gcccagggtc accatccagg tgccttgaggacagagaatg cagaagtggc actgttgaaa 540 tttagaagac catgtgtgaa tggtttcaggcctgggatgt ttgccaccaa gaagtgcctc 600 cgagaaattt ctttcccatt tggaatacagggtggcttga tgggtacggt gggtgaccca 660 acgaagaaaa tgaaattctg ccctttcc 688<210> SEQ ID NO 39 <211> LENGTH: 585 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:14, 15, 24, 53, 108, 135, 465, 477, 495, 499, 504, 517, 530, 580, 581<223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 39 tagtagttgccgcnnaccta aaanttggaa agcatgatgt ctaggaaaca tantaaaata 60 gggtatgcctatgtgctaca gagagatgtt agcatttaaa gtgcatantt ttatgtattt 120 tgacaaatgcatatncctct ataatccaca actgattacg aagctattac aattaaaaag 180 tttggccgggcgtggtgggc ggtggctgac gcctgtaatc ccagcacttt gggaggccga 240 ggcacgcggatcacgaggtc gggagttcaa gaccatcctg gctaacacgg tgaaagtcca 300 tctctactaaaaatacgaaa aaattacccc ggcgtggtgg cgggcgcctg tagtcccagc 360 tactccggaggctgaggcag gagaatggcg tgaacccagg acacggagct tgcagtgtgc 420 caacatcacgtcactgccct ccagcctggg ggacaggaac aagantcccg tcctcanaaa 480 agaaaaatactactnatant ttcnacttta ttttaantta cacagaactn cctcttggta 540 cccccttaccattcatctca cccacctcct atagggcacn nctaa 585 <210> SEQ ID NO 40 <211>LENGTH: 475 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:40 tctgtccaca ccaatcttag aagctctgaa aagaatttgt ctttaaatat cttttaatag 60taacatgtat tttatggacc aaattgacat tttcgactgt tttttccaaa aaagtcaggt 120gaatttcagc acactgagtt gggaatttct tatcccagaa gaccaaccaa tttcatattt 180atttaagatt gattccatac tccgttttca aggagaatcc ctgcagtctc cttaaaggta 240gaacaaatac ttcctatttt tttttcacca ttgtgggatt ggactttaag aggtgactct 300aaaaaaacag agaacaaata tgtctcagtt gtattaagca cggacccata ttatcatatt 360cacttaaaaa aatgatttcc tgtgcacctt ttggcaactt ctcttttcaa tgtagggaaa 420aacttagtca ccctgaaaac ccacaaaata aataaaactt gtagatgtgg acaga 475 <210>SEQ ID NO 41 <211> LENGTH: 423 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 41 taagagggta catcgggtaa gaacgtaggc acatctagagcttagagaag tctggggtag 60 gaaaaaaatc taagtattta taagggtata ggtaacatttaaaagtaggg ctagctgaca 120 ttatttagaa agaacacata cggagagata agggcaaaggactaagacca gaggaacact 180 aatatttagt gatcacttcc attcttggta aaaatagtaacttttaagtt agcttcaagg 240 aagatttttg gccatgatta gttgtcaaaa gttagttctcttgggtttat attactaatt 300 ttgttttaag atccttgtta gtgctttaat aaagtcatgttatatcaaac gctctaaaac 360 attgtagcat gttaaatgtc acaatatact taccatttgttgtatatggc tgtaccctct 420 cta 423 <210> SEQ ID NO 42 <211> LENGTH: 527<212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 470, 475, 515, 522 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 42 tctcctaggc taatgtgtgtgtttctgtaa aagtaaaaag ttaaaaattt taaaaataga 60 aaaaagctta tagaataagaatatgaagaa agaaaatatt tttgtacatt tgcacaatga 120 gtttatgttt taagctaagtgttattacaa aagagccaaa aaggttttaa aaattaaaac 180 gtttgtaaag ttacagtacccttatgttaa tttataattg aagaaagaaa aacttttttt 240 tataaatgta gtgtagcctaagcatacagt atttataaag tctggcagtg ttcaataatg 300 tcctaggcct tcacattcactcactgactc acccagagca acttccagtc ctgtaagctc 360 cattcgtggt aagtgccctatacaggtgca ccatttattt tacagtattt ttactgtacc 420 ttctctatgt ttccatatgtttcgatatac aaataccact ggttactatn gcccnacagg 480 taattccagt aacacggcctgtatacgtct ggtancccta gngaaga 527 <210> SEQ ID NO 43 <211> LENGTH: 331<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 43tcttcaacct cgtaggacaa ctctcatatg cctgggcact atttttaggt tactaccttg 60gctgcccttc tttaagaaaa aaaaaagaag aaaaaagaac ttttccacaa gtttctcttc 120ctctagttgg aaaattagag aaatcatgtt tttaattttg tgttatttca gatcacaaat 180tcaaacactt gtaaacatta agcttctgtt caatcccctg ggaagaggat tcattctgat 240atttacggtt caaaagaagt tgtaatattg tgcttggaac acagagaacc agttattaac 300ttcctactac tattatataa taaataataa c 331 <210> SEQ ID NO 44 <211> LENGTH:592 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 473 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 44 ggcttagtag ttgccaggca aaatarcgttgattctcctc aggagccacc cccaacaccc 60 ctgtttgctt ctagacctat acctagactaaagtcccagc agacccctag aggtgaggtt 120 cagagtgacc cttgaggaga tgtgctacactagaaaagaa ctgcttgagt tttctaattt 180 atataagcag aaatctggag aagagtcataggaatggata ttaagggtgt gagataatgg 240 cggaaggaat atagagttgg atcaggctggacttattgat ttgaacccac taagtagaga 300 ttctgctttt gatgttgcag ctcagggagttaaaaaaggt tttaatggtt ctaatagttt 360 atttgcttgg ttagctgaaa tatggataaaagatggccca ctgtgagcaa gctggaaatg 420 cctgatctct ctcagtttaa tgtagaggaagggatccaaa agtttaggga ganttggatg 480 ctggraktgg attggtcact ttgrgacctacccwtcccag ctgggagggt ccagaagata 540 cacccttgac caacgctttg cgaaatggatttgtgatggc ggcaactact aa 592 <210> SEQ ID NO 45 <211> LENGTH: 567 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 522, 561, 566 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 45 ggcttagtag ttgccattgc gagtgcttgctcaacgagcg ttgaacatgg cggattgtct 60 agattcaacg gatttgagtt ttaccagcaaagcgaaccaa gcgcggccca gagaattatg 120 ggttggttgg ctttgaaaag atggaaatcctgtaggccta gtcagaaaag ccttcttgca 180 gaacagttgg ttctcgggcg aacgctcatcaagatgccca ttggaaaggc tagcgtgtat 240 ttgggagagc ctgatagcgt gtcttctgatgatgtttgtg cttggacagt gacaaaagat 300 atgcaaagca agtccgaact agacgtcaagcttcgtgagc aaattattgt agactcctac 360 ttatactgtg aggaatgata gccaagggtggggactttaa gactaaggtg gtttgtactt 420 gcgccgatga tcccaggcag aaagamctgatcgctagttt tatacgggca actactaagc 480 cgaattccag cacactggcg gccgttactaattggatccg anctcggtac cagcttgatg 540 catascttga gttwtctata ntgtcnc 567<210> SEQ ID NO 46 <211> LENGTH: 908 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:21, 23, 24, 27, 29, 34 <223> OTHER INFORMATION: n = A,T,C or G <400>SEQUENCE: 46 gagcgaaaga ccgagggcag ngnntangng cgangaagcg gagagggccaaaaagcaacc 60 gctttccccg gggggtgccg attcattaag gcaggtggag gacaggtttcccgatggaag 120 gcggcagggg cgcaagcaat taatgtgagt aggccattca ttagcacccgggcttaacat 180 ttaagcttcg ggttggtatg tggtgggaat tgtgagcgga taacaatttcacacaggaaa 240 cagctatgac catgattacg ccaagctatt taggtgacat tatagaataactcaagttat 300 gcatcaagct tggtaccgag ttcggatcca ctagtaacgg ccgccagtgtgtggaattcg 360 gcttagtagt tgccgaccat ggagtgctac ctaggctaga atacctgagytcctccctag 420 cctcactcac attaaattgt atcttttcta cattagatgt cctcagcgccttatttctgc 480 tggacwatcg ataaattaat cctgatagga tgatagcagc agattaattactgagagtat 540 gttaatgtgt catccctcct atataacgta tttgcatttt aatggagcaattctggagat 600 aatccctgaa ggcaaaggaa tgaatcttga gggtgagaaa gccagaatcagtgtccagct 660 gcagttgtgg gagaaggtga tattatgtat gtctcagaag tgacaccatatgggcaacta 720 ctaagcccga attccagcac actggcgggc gttactaatg gatccgagctcggtaccaag 780 cttgatgcat agcttgagta tctatagtgt cactaaatag cctggcgttatcatggtcat 840 agctgtttcc tgtgtgaaat tgttatccgc tcccaattcc ccccaccatacgagccggaa 900 cataaagt 908 <210> SEQ ID NO 47 <211> LENGTH: 480 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 408, 461 <223> OTHER INFORMATION: n = A,T,Cor G <400> SEQUENCE: 47 tgccaacaag gaaagtttta aatttcccct tgaggattcttggtgatcat caaattcagt 60 ggtttttaag gttgttttct gtcaaataac tctaactttaagccaaacag tatatggaag 120 cacagataka atattacaca gataaaagag gagttgatctaaagtaraga tagttggggg 180 ctttaatttc tggaacctag gtctccccat cttcttctgtgctgaggaac ttcttggaag 240 cggggattct aaagttcttt ggaagacagt ttgaaaaccaccatgttgtt ctcagtacct 300 ttatttttaa aaagtaggtg aacattttga gagagaaaagggcttggttg agatgaagtc 360 cccccccccc cttttttttt ttttagctga aatagataccctatgttnaa rgaarggatt 420 attatttacc atgccaytar scacatgctc tttgatgggcnyctccstac cctccttaag 480 <210> SEQ ID NO 48 <211> LENGTH: 591 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 48 aagagggtaccgagtggaat ttccgcttca ctagtctggt gtggctagtc ggtttcgtgg 60 tggccaacattacgaacttc caactcaacc gttcttggac gttcaagcgg gagtaccggc 120 gaggatggtggcgtgaattc tggcctttct ttgccgtggg atcggtagcc gccatcatcg 180 gtatgtttatcaagatcttc tttactaacc cgacctctcc gatttacctg cccgagccgt 240 ggtttaacgaggggaggggg atccagtcac gcgagtactg gtcccagatc ttcgccatcg 300 tcgtgacaatgcctatcaac ttcgtcgtca ataagttgtg gaccttccga acggtgaagc 360 actccgaaaacgtccggtgg ctgctgtgcg gtgactccca aaatcttgat aacaacaagg 420 taaccgaatcgcgctaagga accccggcat ctcgggtact ctgcatatgc gtacccctta 480 agccgaattccagcacactg gcggccgtta ctaattggat ccgaactccg taaccaagcc 540 tgatgcgtaacttgagttat tctatagtgt ccctaaaata acctggcgtt a 591 <210> SEQ ID NO 49<211> LENGTH: 454 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 49 aagagggtac ctgccttgaa atttaaatgt ctaaggaaar tgggagatgattaagagttg 60 gtgtggcyta gtcacaccaa aatgtattta ttacatcctg ctcctttctagttgacagga 120 aagaaagctg ctgtggggaa aggagggata aatactgaag ggatttactaaacaaatgtc 180 catcacagag ttttcctttt tttttttttg agacagagtc ttgctctgtcacccaggctg 240 gaatgaagwg gtatgatctc agttgaatgc aacctctacc tcctaggttcaagcgattct 300 catgcctcag cctcctgagc agctgggact ataggcgcat gctaccatgccaggctaatt 360 tttatatttt tattagagac ggggtgttgc catgttggcc aggcaggtctcgaactcctg 420 ggcctcagat gatctgcccc accgtaccct ctta 454 <210> SEQ ID NO50 <211> LENGTH: 463 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 50 aagagggtac caaaaaaaag aaaaaggaaa aaaagaaaaa caacttgtataaggctttct 60 gctgcataca gctttttttt tttaaataaa tggtgccaac aaatgtttttgcattcacac 120 caattgctgg ttttgaaatc gtactcttca aaggtatttg tgcagatcaatccaatagtg 180 atgccccgta ggttttgtgg actgcccacg ttgtctacct tctcatgtaggagccattga 240 gagactgttt ggacatgcct gtgttcatgt agccgtgatg tccgggggccgtgtacatca 300 tgttaccgtg gggtggggtc tgcattggct gctgggcata tggctgggtgcccatcatgc 360 ccatctgcat ctgcataggg tattggggcg tttgatccat atagccatgattgctgtggt 420 agccactgtt catcattggc tgggacatgc tgttaccctc tta 463 <210>SEQ ID NO 51 <211> LENGTH: 399 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 51 cttcaacctc ccaaagtgct gggattacag gactgagccaccacgctcag cctaagcctc 60 tttttcacta ccctctaagc gatctaccac agtgatgaggggctaaagag cagtgcaatt 120 tgattacaat aatggaactt agatttatta attaacaatttttccttagc atgttggttc 180 cataattatt aagagtatgg acttacttag aaatgagctttcattttaag aatttcatct 240 ttgaccttct ctattagtct gagcagtatg acactatacgtattttattt aactaaccta 300 ccttgagcta ttacttttta aaaggctata tacatgaatgtgtattgtca actgtaaagc 360 cccacagtat ttaattatat catgatgtct ttgaggttg 399<210> SEQ ID NO 52 <211> LENGTH: 392 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 52 cttcaacctc aatcaacctt ggtaattgataaaatcatca cttaactttc tgatataatg 60 gcaataatta tctgagaaaa aaaagtggtgaaagattaaa cttgcatttc tctcagaatc 120 ttgaaggata tttgaataat tcaaaagcggaatcagtagt atcagccgaa gaaactcact 180 tagctagaac gttggaccca tggatctaagtccctgccct tccactaacc agctgattgg 240 ttttgtgtaa acctcctaca cgcttgggcttggtcgcctc atttgtcaaa gtaaaggctg 300 aaataggaag ataatgaacc gtgtctttttggtctctttt ccatccatta ctctgatttt 360 acaaagaggc ctgtattccc ctggtgaggt tg392 <210> SEQ ID NO 53 <211> LENGTH: 179 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:135, 143, 179 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 53ttcgggtgat gcctcctcag gctacagtga agactggatt acagaaaggt gccagcgaga 60tttcagattc ctgtaaacct ctaaagaaaa ggagtcgcgc ctcaactgat gtagaaatga 120ctagttcagc atacngagac acntctgact ccgattctag aggactgagt gacctgcan 179<210> SEQ ID NO 54 <211> LENGTH: 112 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:31, 49, 54, 55, 75, 91, 107 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 54 ttcgggtgat gcctcctcag gctacatcat natagaagcaaagtagaana atcnngtttg 60 tgcattttcc cacanacaaa attcaaatga ntggaagaaattggganagt at 112 <210> SEQ ID NO 55 <211> LENGTH: 225 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 55 tgagcttccg cttctgacaactcaatagat aatcaaagga caactttaac agggattcac 60 aaaggagtat atccaaatgccaataaacat ataaaaagga attcagcttc atcatcatca 120 gaagwatgca aattaaaaccataatgagaa accactatgt cccactagaa tagataaaat 180 cttaaaagac tggtaaaaccaagtgttggt aaggcaagag gagca 225 <210> SEQ ID NO 56 <211> LENGTH: 175<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 56gctcctcttg ccttaccaac acattctcaa aaacctgtta gagtcctaag cattctcctg 60ttagtattgg gattttaccc ctgtcctata aagatgttat gtaccaaaaa tgaagtggag 120ggccataccc tgagggaggg gagggatctc tagtgttgtc agaagcggaa gctca 175 <210>SEQ ID NO 57 <211> LENGTH: 223 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 57 agccatttac cacccatgga tgaatggatt ttgtaattctagctgttgta ttttgtgaat 60 ttgttaattt tgttgttttt ctgtgaaaca catacattggatatgggagg taaaggagtg 120 tcccagttgc tcctggtcac tccctttata gccattactgtcttgtttct tgtaactcag 180 gttaggtttt ggtctctctt gctccactgc aaaaaaaaaaaaa 223 <210> SEQ ID NO 58 <211> LENGTH: 211 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 58 gttcgaaggt gaacgtgtaggtagcggatc tcacaactgg ggaactgtca aagacgaatt 60 aactgacttg gatcaatcaaatgtgactga ggaaacacct gaaggtgaag aacatcatcc 120 agtggcagac actgaaaataaggagaatga agttgaagag gtaaaagagg agggtccaaa 180 agagatgact ttggatgggtggtaaatggc t 211 <210> SEQ ID NO 59 <211> LENGTH: 208 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 59 gctcctcttg ccttaccaactttgcaccca tcatcaacca tgtggccagg tttgcagccc 60 aggctgcaca tcaggggactgcctcgcaat acttcatgct gttgctgctg actgatggtg 120 ctgtgacgga tgtggaagccacacgtgagg ctgtggtgcg tgcctcgaac ctgcccatgt 180 cagtgatcat tatgggtggtaaatggct 208 <210> SEQ ID NO 60 <211> LENGTH: 171 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 60 agccatttac cacccatactaaattctagt tcaaactcca acttcttcca taaaacatct 60 aaccactgac accagttggcaatagcttct tccttcttta acctcttaga gtatttatgg 120 tcaatgccac acatttctgcaactgaataa agttggtaag gcaagaggag c 171 <210> SEQ ID NO 61 <211> LENGTH:134 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 37, 70, 80, 86, 88, 97, 117, 123,131 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 61cgggtgatgc ctcctcaggc tttggtgtgt ccactcnact cactggcctc ttctccagca 60actggtgaan atgtcctcan gaaaancncc acacgcngct cagggtgggg tgggaancat 120canaatcatc nggc 134 <210> SEQ ID NO 62 <211> LENGTH: 145 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 62 agagggtaca tatgcaacagtatataaagg aagaagtgca ctgagaggaa cttcatcaag 60 gccatttaat caataagtgatagagtcaag gctcaaccca ggtgtgacgg attccaggtc 120 ccaagctcct tactggtaccctctt 145 <210> SEQ ID NO 63 <211> LENGTH: 297 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 63 tgcactgaga ggaattcaaagggtttatgc caaagaacaa accagtcctc tgcagcctaa 60 ctcatttgtt tttgggctgcgaagccatgt agagggcgat caggcagtag atggtccctc 120 ccacagtcag cgccatggtggtccggtaaa gcatttggtc aggcaggcct cgtttcaggt 180 agacgggcac acatcagctttctggaaaaa cttttgtagc tctggagctt tgtttttccc 240 agcataatca tacactgtggaatcggaggt cagtttagtt ggtaaggcaa gaggagc 297 <210> SEQ ID NO 64 <211>LENGTH: 300 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:64 gcactgagag gaacttccaa tactatgttg aataggagtg gtgagagagg gcatccttgt 60cttgtgccgg ttttcaaagg gaatgcttcc agcttttgcc cattcagtat aatattaaag 120aatgttttac cattttctgt cttgcctgtt tttctgtgtt tttgttggtc tcttcattct 180ccatttttag gcctttacat gttaggaata tatttctttt aatgatactt cacctttggt 240atcttttgtg agactctact catagtgtga taagcactgg gttggtaagg caagaggagc 300<210> SEQ ID NO 65 <211> LENGTH: 203 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 65 gctcctcttg ccttaccaac tcacccagtatgtcagcaat tttatcrgct ttacctacga 60 aacagcctgt atccaaacac ttaacacactcacctgaaaa gttcaggcaa caatcgcctt 120 ctcatgggtc tctctgctcc agttctgaacctttctcttt tcctagaaca tgcatttarg 180 tcgatagaag ttcctctcag tgc 203 <210>SEQ ID NO 66 <211> LENGTH: 344 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 66 tacggggacc cctgcattga gaaagcgaga ctcactctgaagctgaaatg ctgttgccct 60 tgcagtgctg gtagcaggag ttctgtgctt tgtgggctaaggctcctgga tgacccctga 120 catggagaag gcagagttgt gtgccccttc tcatggcctcgtcaaggcat catggactgc 180 cacacacaaa atgccgtttt tattaacgac atgaaattgaaggagagaac acaattcact 240 gatgtggctc gtaaccatgg atatggtcac atacagaggtgtgattatgt aaaggttaat 300 tccacccacc tcatgtggaa actagcctca atgcaggggtccca 344 <210> SEQ ID NO 67 <211> LENGTH: 157 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 67 gcactgagag gaacttcgtagggaggttga actggctgct gaggaggggg aacaacaggg 60 taaccagact gatagccattggatggataa tatggtggtt gaggagggac actacttata 120 gcagagggtt gtgtatagcctgaggaggca tcacccg 157 <210> SEQ ID NO 68 <211> LENGTH: 137 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 68 gcactgagaggaacttctag aaagtgaaag tctagacata aaataaaata aaaatttaaa 60 actcaggagagacagcccag cacggtggct cacgcctgta atcccagaac tttgggagcc 120 tgaggaggcatcacccg 137 <210> SEQ ID NO 69 <211> LENGTH: 137 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 69 cgggtgatgc ctcctcaggctgtattttga agactatcga ctggacttct tatcaactga 60 agaatccgtt aaaaataccagttgtattat ttctacctgt caaaatccat ttcaaatgtt 120 gaagttcctc tcagtgc 137<210> SEQ ID NO 70 <211> LENGTH: 220 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:89, 112, 129, 171, 172 <223> OTHER INFORMATION: n = A,T,C or G <400>SEQUENCE: 70 agcatgttga gcccagacac gcaatctgaa tgagtgtgca cctcaagtaaatgtctacac 60 gctgcctggt ctgacatggc acaccatcnc gtggagggca casctctgctcngcctacwa 120 cgagggcant ctcatwgaca ggttccaccc accaaactgc aagaggctcannaagtactr 180 ccagggtmya sggacmasgg tgggaytyca ycacwcatct 220 <210> SEQID NO 71 <211> LENGTH: 353 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 66, 160,204, 246, 267, 334, 339, 342 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 71 cgttagggtc tctatccact gctaaaccat acacctgggtaaacagggac catttaacat 60 tcccanctaa atatgccaag tgacttcaca tgtttatcttaaagatgtcc aaaacgcaac 120 tgattttctc ccctaaacct gtgatggtgg gatgattaancctgagtggt ctacagcaag 180 ttaagtgcaa ggtgctaaat gaangtgacc tgagatacagcatctacaag gcagtacctc 240 tcaacncagg gcaactttgc ttctcanagg gcatttagcagtgtctgaag taatttctgt 300 attacaactc acggggcggg gggtgaatat ctantgganagnagacccta acg 353 <210> SEQ ID NO 72 <211> LENGTH: 343 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 72 gcactgagag gaacttccaatacyatkatc agagtgaaca rgcarccyac agaacaggag 60 aaaatgttyg caatctctccatctgacaaa aggctaatat ccagawtcta awaggaactt 120 aaacaaattt atgagaaaagaacaracaac ctcawcaaaa agtgggtgaa ggawatgcts 180 aaargaagac atytattcagccagtaaaca yatgaaaaaa aggctcatsa tcactgawca 240 ttagagaaat gcaaatcaaaaccacaatga gataccatct yayrccagtt agaayggtga 300 tcattaaaar stcaggaaacaacagatgct ggacaaggtg tca 343 <210> SEQ ID NO 73 <211> LENGTH: 321 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 288 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 73 gcactgagag gaacttcaga gagagagaga gagttccaccctgtacttgg ggagagaaac 60 agaaggtgag aaagtctttg gttctgaagc agcttctaagatcttttcat ttgcttcatt 120 tcaaagttcc catgctgcca aagtgccatc ctttggggtactgttttctg agctccagtg 180 ataactcatt tatacaaggg agatacccag aaaaaaagtgagcaaatctt aaaaaggtgg 240 cttgagttca gccttaaata ccatcttgaa atgacacagagaaagaanga tgttgggtgg 300 gagtggatag agaccctaac g 321 <210> SEQ ID NO 74<211> LENGTH: 321 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 74 gcactgagag gaacttcaga gagagagaga gagttccacc ctgtacttggggagagaaac 60 agaaggtgag aaagtctttg gttctgaagc agcttctaag atcttttcatttgcttcatt 120 tcaaagttcc catgctgcca aagtgccatc ctttggggta ctgttttctgagctccagtg 180 ataactcatt tatacaaggg agatacccag aaaaaaagtg agcaaatcttaaaaaggtgg 240 cttgagttca gycttaaata ccatcttgaa atgamacaga gaaagaaggatgttgggtgg 300 gagtggatag agaccctaac g 321 <210> SEQ ID NO 75 <211>LENGTH: 317 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:75 gcactgagag gaacttccac atgcactgag aaatgcatgt tcacaaggac tgaagtctgg 60aactcagttt ctcagttcca atcctgattc aggtgtttac cagctacaca accttaagca 120agtcagataa ccttagcttc ctcatatgca aaatgagaat gaaaagtact catcgctgaa 180ttgttttgag gattagaaaa acatctggca tgcagtagaa attcaattag tattcatttt 240cattcttcta aattaaacaa ataggatttt tagtggtgga acttcagaca ccagaaatgg 300gagtggatag agaccct 317 <210> SEQ ID NO 76 <211> LENGTH: 244 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 76 cgttagggtctctatccact cccactactg atcaaactct atttatttaa ttatttttat 60 catactttaagttctgggat acacgtgcag catgcgcagg tttgttgcat aggtatacac 120 ttgccatggtggtttgctgc acccatcagt ccatcatcta cattaggtat ttctcctaat 180 gctatccctcccctagcccc ttacaccccc aacaggctct agtgtgtgaa gttcctctca 240 gtgc 244<210> SEQ ID NO 77 <211> LENGTH: 254 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 77 cgttagggtc tctatccact gaaatctgaagcacaggagg aagagaagca gtyctagtga 60 gatggcaagt tcwtttacca cactctttaacatttygttt agttttaacc tttatttatg 120 gataataaag gttaatatta ataatgatttattttaaggc attcccraat ttgcataatt 180 ctccttttgg agataccctt ttatctccagtgcaagtctg gatcaaagtg atasamagaa 240 gttcctctca gtgc 254 <210> SEQ ID NO78 <211> LENGTH: 355 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 69, 87, 186, 192,220, 227, 251, 278, 339, 346, 350 <223> OTHER INFORMATION: n = A,T,C orG <400> SEQUENCE: 78 ttcgatacag gcaaacatga actgcaggag ggtggtgacgatcatgatgt tgccgatggt 60 ccggatggnc acgaagacgc actggancac gtgcttacgtccttttgctc tgttgatggc 120 cctgagggga cgcaggaccc ttatgaccct cagaatcttcacaacgggag atggcactgg 180 attgantccc antgacacca gagacacccc aaccaccagnatatcantat attgatgtag 240 ttcctgtaga nggccccctt gtggaggaaa gctccatnagttggtcatct tcaacaggat 300 ctcaacagtt tccgatggct gtgatgggca tagtcatanttaaccntgtn tcgaa 355 <210> SEQ ID NO 79 <211> LENGTH: 406 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 79 taagagggtaccagcagaaa ggttagtatc atcagatagc atcttatacg agtaatatgc 60 ctgctatttgaagtgtaatt gagaaggaaa attttagcgt gctcactgac ctgcctgtag 120 ccccagtgacagctaggatg tgcattctcc agccatcaag agactgagtc aagttgttcc 180 ttaagtcagaacagcagact cagctctgac attctgattc gaatgacact gttcaggaat 240 cggaatcctgtcgattagac tggacagctt gtggcaagtg aatttgcctg taacaagcca 300 gattttttaaaatttatatt gtaaataatg tgtgtgtgtg tgtgtgtata tatatatata 360 tgtacagttatctaagttaa tttaaaagtt gtttggtacc ctctta 406 <210> SEQ ID NO 80 <211>LENGTH: 327 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:80 tttttttttt tttactcggc tcagtctaat cctttttgta gtcactcata ggccagactt 60agggctagga tgatgattaa taagagggat gacataacta ttagtggcag gttagttgtt 120tgtagggctc atggtagggg taaaaggagg gcaatttcta gatcaaataa taagaaggta 180atagctacta agaagaattt tatggagaaa gggacgcggg cgggggatat agggtcgaag 240ccgcactcgt aaggggtgga tttttctatg tagccgttga gttgtggtag tcaaaatgta 300ataattatta gtagtaagcc taggaga 327 <210> SEQ ID NO 81 <211> LENGTH: 318<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 81tagtctatgc ggttgattcg gcaatccatt atttgctgga ttttgtcatg tgttttgcca 60attgcattca taatttatta tgcatttatg cttgtatctc ctaagtcatg gtatataatc 120catgcttttt atgttttgtc tgacataaac tcttatcaga gccctttgca cacagggatt 180caataaatat taacacagtc tacatttatt tggtgaatat tgcatatctg ctgtactgaa 240agcacattaa gtaacaaagg caagtgagaa gaatgaaaag cactactcac aacagttatc 300atgattgcgc atagacta 318 <210> SEQ ID NO 82 <211> LENGTH: 338 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 82 tcttcaacctctactcccac taatagcttt ttgatgactt ctagcaagcc tcgctaacct 60 cgccttaccccccactatta acctactggg agaactctct gtgctagtaa ccacgttctc 120 ctgatcaaatatcactctcc tacttacagg actcaacata ctagtcacag ccctatactc 180 cctctacatatttaccacaa cacaatgggg ctcactcacc caccacatta acaacataaa 240 accctcattcacacgagaaa acaccctcat gttcatacac ctatccccca ttctcctcct 300 atccctcaaccccgacatca ttaccgggtt ttcctctt 338 <210> SEQ ID NO 83 <211> LENGTH: 111<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 83agccatttac cacccatcca caaaaaaaaa aaaaaaaaag aaaaatatca aggaataaaa 60atagactttg aacaaaaagg aacatttgct ggcctgagga ggcatcaccc g 111 <210> SEQID NO 84 <211> LENGTH: 224 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 84 tcgggtgatg cctcctcagg ccaagaagat aaagcttcagacccctaaca catttccaaa 60 aaggaagaaa ggagaaaaaa gggcatcatc cccgttccgaagggtcaggg aggaggaaat 120 tgaggtggat tcacgagttg cggacaactc ctttgatgccaagcgaggtg cagccggaga 180 ctggggagag cgagccaatc aggttttgaa gttcctctcagtgc 224 <210> SEQ ID NO 85 <211> LENGTH: 348 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 85 gcactgagag gaacttcgttggaaacgggt ttttttcatg taaggctaga cagaagaatt 60 ctcagtaact tccttgtgttgtgtgtattc aactcacasa gttgaacgat cctttacaca 120 gagcagactt gtaacactcttwttgtggaa tttgcaagtg gagatttcag scgctttgaa 180 gtsaaaggta gaaaaggaaatatcttccta taaaaactag acagaatgat tctcagaaac 240 tcctttgtga tgtgtgcgttcaactcacag agtttaacct ttcwtttcat agaagcagtt 300 aggaaacact ctgtttgtaaagtctgcaag tggatagaga ccctaacg 348 <210> SEQ ID NO 86 <211> LENGTH: 293<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 86gcactgagag gaacttcytt gtgwtgtktg yattcaactc acagagttga asswtsmttt 60acabagwkca ggcttkcaaa cactcttttt gtmgaatytg caagwggaka tttsrrccrc 120tttgwggycw wysktmgaaw mggrwatatc ttcwyatmra amctagacag aaksattctc 180akaawstyyy ytgtgawgws tgcrttcaac tcacagagkt kaacmwtyct kytsatrgag 240cagttwkgaa actctmtttc tttggattct gcaagtggat agagacccta acg 293 <210> SEQID NO 87 <211> LENGTH: 10 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer foramplification from breast cancer tumor cDNA <400> SEQUENCE: 87ctcctaggct 10 <210> SEQ ID NO 88 <211> LENGTH: 10 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:PCR primer for amplification from breast cancer tumor cDNA <400>SEQUENCE: 88 agtagttgcc 10 <210> SEQ ID NO 89 <211> LENGTH: 11 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: PCR primer for amplification from breast cancer tumor cDNA<400> SEQUENCE: 89 ttccgttatg c 11 <210> SEQ ID NO 90 <211> LENGTH: 10<212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223>OTHER INFORMATION: PCR primer for amplification from breast cancer tumorcDNA <400> SEQUENCE: 90 tggtaaaggg 10 <210> SEQ ID NO 91 <211> LENGTH:10 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: PCR primer for amplification from breast cancertumor cDNA <400> SEQUENCE: 91 tcggtcatag 10 <210> SEQ ID NO 92 <211>LENGTH: 10 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: PCR primer for amplification frombreast cancer tumor cDNA <400> SEQUENCE: 92 tacaacgagg 10 <210> SEQ IDNO 93 <211> LENGTH: 10 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer foramplification from breast cancer tumor cDNA <400> SEQUENCE: 93tggattggtc 10 <210> SEQ ID NO 94 <211> LENGTH: 10 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:PCR primer for amplification from breast cancer tumor cDNA <400>SEQUENCE: 94 ctttctaccc 10 <210> SEQ ID NO 95 <211> LENGTH: 10 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: PCR primer for amplification from breast cancer tumor cDNA<400> SEQUENCE: 95 ttttggctcc 10 <210> SEQ ID NO 96 <211> LENGTH: 10<212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223>OTHER INFORMATION: PCR primer for amplification from breast cancer tumorcDNA <400> SEQUENCE: 96 ggaaccaatc 10 <210> SEQ ID NO 97 <211> LENGTH:10 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: PCR primer for amplification from breast cancertumor cDNA <400> SEQUENCE: 97 tcgatacagg 10 <210> SEQ ID NO 98 <211>LENGTH: 10 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: PCR primer for amplification frombreast cancer tumor cDNA <400> SEQUENCE: 98 ggtactaagg 10 <210> SEQ IDNO 99 <211> LENGTH: 10 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer foramplification from breast cancer tumor cDNA <400> SEQUENCE: 99agtctatgcg 10 <210> SEQ ID NO 100 <211> LENGTH: 10 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:PCR primer for amplification from breast cancer tumor cDNA <400>SEQUENCE: 100 ctatccatgg 10 <210> SEQ ID NO 101 <211> LENGTH: 10 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: PCR primer for amplification from breast cancer tumor cDNA<400> SEQUENCE: 101 tctgtccaca 10 <210> SEQ ID NO 102 <211> LENGTH: 10<212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223>OTHER INFORMATION: PCR primer for amplification from breast cancer tumorcDNA <400> SEQUENCE: 102 aagagggtac 10 <210> SEQ ID NO 103 <211> LENGTH:10 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: PCR primer for amplification from breast cancertumor cDNA <400> SEQUENCE: 103 cttcaacctc 10 <210> SEQ ID NO 104 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: PCR primer for amplification frombreast cancer tumor cDNA <400> SEQUENCE: 104 gctcctcttg ccttaccaac 20<210> SEQ ID NO 105 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primerfor amplification from breast cancer tumor cDNA <400> SEQUENCE: 105gtaagtcgag cagtgtgatg 20 <210> SEQ ID NO 106 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: PCR primer for amplification from breast cancer tumor cDNA<400> SEQUENCE: 106 gtaagtcgag cagtctgatg 20 <210> SEQ ID NO 107 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: PCR primer for amplification frombreast cancer tumor cDNA <400> SEQUENCE: 107 gacttagtgg aaagaatgta 20<210> SEQ ID NO 108 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primerfor amplification from breast cancer tumor cDNA <400> SEQUENCE: 108gtaattccgc caaccgtagt 20 <210> SEQ ID NO 109 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: PCR primer for amplification from breast cancer tumor cDNA<400> SEQUENCE: 109 atggttgatc gatagtggaa 20 <210> SEQ ID NO 110 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: PCR primer for amplification frombreast cancer tumor cDNA <400> SEQUENCE: 110 acggggaccc ctgcattgag 20<210> SEQ ID NO 111 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primerfor amplification from breast cancer tumor cDNA <400> SEQUENCE: 111tattctagac cattcgctac 20 <210> SEQ ID NO 112 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: PCR primer for amplification from breast cancer tumor cDNA<400> SEQUENCE: 112 acataaccac tttagcgttc 20 <210> SEQ ID NO 113 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: PCR primer for amplification frombreast cancer tumor cDNA <400> SEQUENCE: 113 cgggtgatgc ctcctcaggc 20<210> SEQ ID NO 114 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primerfor amplification from breast cancer tumor cDNA <400> SEQUENCE: 114agcatgttga gcccagacac 20 <210> SEQ ID NO 115 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: PCR primer for amplification from breast cancer tumor cDNA<400> SEQUENCE: 115 gacaccttgt ccagcatctg 20 <210> SEQ ID NO 116 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: PCR primer for amplification frombreast cancer tumor cDNA <400> SEQUENCE: 116 tacgctgcaa cactgtggag 20<210> SEQ ID NO 117 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primerfor amplification from breast cancer tumor cDNA <400> SEQUENCE: 117cgttagggtc tctatccact 20 <210> SEQ ID NO 118 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: PCR primer for amplification from breast cancer tumor cDNA<400> SEQUENCE: 118 agactgactc atgtccccta 20 <210> SEQ ID NO 119 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: PCR primer for amplification frombreast cancer tumor cDNA <400> SEQUENCE: 119 tcatcgctcg gtgactcaag 20<210> SEQ ID NO 120 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primerfor amplification from breast cancer tumor cDNA <400> SEQUENCE: 120caagattcca taggctgacc 20 <210> SEQ ID NO 121 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: PCR primer for amplification from breast cancer tumor cDNA<400> SEQUENCE: 121 acgtactggt cttgaaggtc 20 <210> SEQ ID NO 122 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: PCR primer for amplification frombreast cancer tumor cDNA <400> SEQUENCE: 122 gacgcttggc cacttgacac 20<210> SEQ ID NO 123 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primerfor amplification from breast cancer tumor cDNA <400> SEQUENCE: 123gtatcgacgt agtggtctcc 20 <210> SEQ ID NO 124 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: PCR primer for amplification from breast cancer tumor cDNA<400> SEQUENCE: 124 tagtgacatt acgacgctgg 20 <210> SEQ ID NO 125 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: PCR primer for amplification frombreast cancer tumor cDNA <400> SEQUENCE: 125 cgggtgatgc ctcctcaggc 20<210> SEQ ID NO 126 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primerfor amplification from breast cancer tumor cDNA <400> SEQUENCE: 126atggctattt tcgggggctg aca 23 <210> SEQ ID NO 127 <211> LENGTH: 22 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: PCR primer for amplification from breast cancer tumor cDNA<400> SEQUENCE: 127 ccggtatctc ctcgtgggta tt 22 <210> SEQ ID NO 128<211> LENGTH: 18 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: PCR primer for amplificationfrom breast cancer tumor cDNA <400> SEQUENCE: 128 ctgcctgagc cacaaatg 18<210> SEQ ID NO 129 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primerfor amplification from breast cancer tumor cDNA <400> SEQUENCE: 129ccggaggagg aagctagagg aata 24 <210> SEQ ID NO 130 <211> LENGTH: 14 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: PCR primer for amplification from breast cancer tumor cDNA<400> SEQUENCE: 130 tttttttttt ttag 14 <210> SEQ ID NO 131 <211> LENGTH:18 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Predicited Th Motifs (B-cell epitopes) <400>SEQUENCE: 131 Ser Ser Gly Gly Arg Thr Phe Asp Asp Phe His Arg Tyr LeuLeu Val 1 5 10 15 Gly Ile <210> SEQ ID NO 132 <211> LENGTH: 22 <212>TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Predicited Th Motifs (B-cell epitopes) <220> FEATURE: <221>NAME/KEY: VARIANT <222> LOCATION: 13 <223> OTHER INFORMATION: Xaa = AnyAmino Acid <400> SEQUENCE: 132 Gln Gly Ala Ala Gln Lys Pro Ile Asn LeuSer Lys Xaa Ile Glu Val 1 5 10 15 Val Gln Gly His Asp Glu 20 <210> SEQID NO 133 <211> LENGTH: 23 <212> TYPE: PRT <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Predicited Th Motifs(B-cell epitopes) <400> SEQUENCE: 133 Ser Pro Gly Val Phe Leu Glu HisLeu Gln Glu Ala Tyr Arg Ile Tyr 1 5 10 15 Thr Pro Phe Asp Leu Ser Ala 20<210> SEQ ID NO 134 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PredicitedHLA A2.1 Motifs (T-cell epitopes) <400> SEQUENCE: 134 Tyr Leu Leu ValGly Ile Gln Gly Ala 1 5 <210> SEQ ID NO 135 <211> LENGTH: 9 <212> TYPE:PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Predicited HLA A2.1 Motifs (T-cell epitopes) <400>SEQUENCE: 135 Gly Ala Ala Gln Lys Pro Ile Asn Leu 1 5 <210> SEQ ID NO136 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Predicited HLA A2.1 Motifs(T-cell epitopes) <220> FEATURE: <221> NAME/KEY: VARIANT <222> LOCATION:5 <223> OTHER INFORMATION: Xaa = Any Amino Acid <400> SEQUENCE: 136 AsnLeu Ser Lys Xaa Ile Glu Val Val 1 5 <210> SEQ ID NO 137 <211> LENGTH: 9<212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223>OTHER INFORMATION: Predicited HLA A2.1 Motifs (T-cell epitopes) <400>SEQUENCE: 137 Glu Val Val Gln Gly His Asp Glu Ser 1 5 <210> SEQ ID NO138 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Predicited HLA A2.1 Motifs(T-cell epitopes) <400> SEQUENCE: 138 His Leu Gln Glu Ala Tyr Arg IleTyr 1 5 <210> SEQ ID NO 139 <211> LENGTH: 9 <212> TYPE: PRT <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Predicited HLA A2.1 Motifs (T-cell epitopes) <400> SEQUENCE: 139 Asn LeuAla Phe Val Ala Gln Ala Ala 1 5 <210> SEQ ID NO 140 <211> LENGTH: 9<212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223>OTHER INFORMATION: Predicited HLA A2.1 Motifs (T-cell epitopes) <400>SEQUENCE: 140 Phe Val Ala Gln Ala Ala Pro Asp Ser 1 5 <210> SEQ ID NO141 <211> LENGTH: 9388 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 141 gctcgcggcc gcgagctcaa ttaaccctca ctaaagggagtcgactcgat cagactgtta 60 ctgtgtctat gtagaaagaa gtagacataa gagattccattttgttctgt actaagaaaa 120 attcttctgc cttgagatgc tgttaatctg taaccctagccccaaccctg tgctcacaga 180 gacatgtgct gtgttgactc aaggttcaat ggatttagggctatgctttg ttaaaaaagt 240 gcttgaagat aatatgcttg ttaaaagtca tcaccattctctaatctcaa gtacccaggg 300 acacaataca ctgcggaagg ccgcagggac ctctgtctaggaaagccagg tattgtccaa 360 gatttctccc catgtgatag cctgagatat ggcctcatgggaagggtaag acctgactgt 420 cccccagccc gacatccccc agcccgacat cccccagcccgacacccgaa aagggtctgt 480 gctgaggagg attagtaaaa gaggaaggcc tctttgcagttgaggtaaga ggaaggcatc 540 tgtctcctgc tcgtccctgg gcaatagaat gtcttggtgtaaaacccgat tgtatgttct 600 acttactgag ataggagaaa acatccttag ggctggaggtgagacacgct ggcggcaata 660 ctgctcttta atgcaccgag atgtttgtat aagtgcacatcaaggcacag cacctttcct 720 taaacttatt tatgacacag agacctttgt tcacgttttcctgctgaccc tctccccact 780 attaccctat tggcctgcca catccccctc tccgagatggtagagataat gatcaataaa 840 tactgaggga actcagagac cagtgtccct gtaggtcctccgtgtgctga gcgccggtcc 900 cttgggctca cttttctttc tctatacttt gtctctgtgtctctttcttt tctcagtctc 960 tcgttccacc tgacgagaaa tacccacagg tgtggaggggcaggccaccc cttcaataat 1020 ttactagcct gttcgctgac aacaagactg gtggtgcagaaggttgggtc ttggtgttca 1080 ccgggtggca ggcatgggcc aggtgggagg gtctccagcgcctggtgcaa atctccaaga 1140 aagtgcagga aacagcacca agggtgattg taaattttgatttggcgcgg caggtagcca 1200 ttccagcgca aaaatgcgca ggaaagcttt tgctgtgcttgtaggcaggt aggccccaag 1260 cacttcttat tggctaatgt ggagggaacc tgcacatccattggctgaaa tctccgtcta 1320 tttgaggctg actgagcgcg ttcctttctt ctgtgttgcctggaaacgga ctgtctgcct 1380 agtaacatct gatcacgttt cccattggcc gccgtttccggaagcccgcc ctcccatttc 1440 cggaagcctg gcgcaaggtt ggtctgcagg tggcctccaggtgcaaagtg ggaagtgtga 1500 gtcctcagtc ttgggctatt cggccacgtg cctgccggacatgggacgct ggagggtcag 1560 cagcgtggag tcctggcctt ttgcgtccac gggtgggaaattggccattg ccacggcggg 1620 aactgggact caggctgccc cccggccgtt tctcatccgtccaccggact cgtgggcgct 1680 cgcactggcg ctgatgtagt ttcctgacct ctgacccgtattgtctccag attaaaggta 1740 aaaacggggc tttttcagcc cactcgggta aaacgccttttgatttctag gcaggtgttt 1800 tgttgcacgc ctgggaggga gtgacccgca ggttgaggtttattaaaata cattcctggt 1860 ttatgttatg tttataataa agcaccccaa cctttacaaaatctcacttt ttgccagttg 1920 tattatttag tggactgtct ctgataagga cagccagttaaaatggaatt ttgttgttgc 1980 taattaaacc aatttttagt tttggtgttt gtcctaatagcaacaacttc tcaggcttta 2040 taaaaccata tttcttgggg gaaatttctg tgtaaggcacagcgagttag tttggaattg 2100 ttttaaagga agtaagttcc tggttttgat atcttagtagtgtaatgccc aacctggttt 2160 ttactaaccc tgtttttaga ctctcccttt ccttaaatcacctagccttg tttccacctg 2220 aattgactct cccttagcta agagcgccag atggactccatcttggctct ttcactggca 2280 gccccttcct caaggactta acttgtgcaa gctgactcccagcacatcca agaatgcaat 2340 taactgttaa gatactgtgg caagctatat ccgcagttccgaggaattca tccgattgat 2400 tatgcccaaa agccccgcgt ctatcacctt gtaataatcttaaagcccct gcacctggaa 2460 ctattaactt tcctgtaacc atttatcctt ttaacttttttgcttacttt atttctgtaa 2520 aattgtttta actagacctc ccctcccctt tctaaaccaaagtataaaag aagatctagc 2580 cccttcttca gagcggagag aattttgagc attagccatctcttggcggc cagctaaata 2640 aatggacttt taatttgtct caaagtgtgg cgttttctctaactcgctca ggtacgacat 2700 ttggaggccc cagcgagaaa cgtcaccggg agaaacgtcaccgggcgaga gccgggcccg 2760 ctgtgtgctc ccccggaagg acagccagct tgtaggggggagtgccacct gaaaaaaaaa 2820 tttccaggtc cccaaagggt gaccgtcttc cggaggacagcggatcgact accatgcggg 2880 tgcccaccaa aattccacct ctgagtcctc aactgctgaccccggggtca ggtaggtcag 2940 atttgacttt ggttctggca gagggaagcg accctgatgagggtgtccct cttttgactc 3000 tgcccatttc tctaggatgc tagagggtag agccctggttttctgttaga cgcctctgtg 3060 tctctgtctg ggagggaagt ggccctgaca ggggccatcccttgagtcag tccacatccc 3120 aggatgctgg gggactgagt cctggtttct ggcagactggtctctctctc tctctttttc 3180 tatctctaat ctttccttgt tcaggtttct tggagaatctctgggaaaga aaaaagaaaa 3240 actgttataa actctgtgtg aatggtgaat gaatgggggaggacaagggc ttgcgcttgt 3300 cctccagttt gtagctccac ggcgaaagct acggagttcaagtgggccct cacctgcggt 3360 tccgtggcga cctcataagg cttaaggcag catccggcatagctcgatcc gagccggggg 3420 tttataccgg cctgtcaatg ctaagaggag cccaagtcccctaaggggga gcggccaggc 3480 gggcatctga ctgatcccat cacgggaccc cctccccttgtttgtctaaa aaaaaaaaaa 3540 gaagaaactg tcataactgt ttacatgccc tagggtcaactgtttgtttt atgtttattg 3600 ttctgttcgg tgtctattgt cttgtttagt ggttgtcaaggttttgcatg tcaggacgtc 3660 gatattgccc aagacgtctg ggtaagaact tctgcaaggtccttagtgct gattttttgt 3720 cacaggaggt taaatttctc atcaatcatt taggctggccaccacagtcc tgtcttttct 3780 gccagaagca agtcaggtgt tgttacggga atgagtgtaaaaaaacattc gcctgattgg 3840 gatttctggc accatgatgg ttgtatttag attgtcataccccacatcca ggttgattgg 3900 acctcctcta aactaaactg gtggtgggtt caaaacagccaccctgcaga tttccttgct 3960 cacctctttg gtcattctgt aacttttcct gtgcccttaaatagcacact gtgtagggaa 4020 acctaccctc gtactgcttt acttcgttta gattcttactctgttcctct gtggctactc 4080 tcccatctta aaaacgatcc aagtggtcct tttcctcctccctgccccct accccacaca 4140 tctcgttttc cagtgcgaca gcaagttcag cgtctccaggacttggctct gctctcactc 4200 cttgaaccct taaaagaaaa agctgggttt gagctatttgcctttgagtc atggagacac 4260 aaaaggtatt tagggtacag atctagaaga agagagagaacacctagatc caactgaccc 4320 aggagatctc gggctggcct ctagtcctcc tccctcaatcttaaagctac agtgatgtgg 4380 caagtggtat ttagctgttg tggtttttct gctctttctggtcatgttga ttctgttctt 4440 tcgatactcc agccccccag ggagtgagtt tctctgtctgtgctgggttt gatatctatg 4500 ttcaaatctt attaaattgc cttcaaaaaa aaaaaaaaaagggaaacact tcctcccagc 4560 cttgtaaggg ttggagccct ctccagtata tgctgcagaatttttctctc ggtttctcag 4620 aggattatgg agtccgcctt aaaaaaggca agctctggacactctgcaaa gtagaatggc 4680 caaagtttgg agttgagtgg ccccttgaag ggtcactgaacctcacaatt gttcaagctg 4740 tgtggcgggt tgttactgaa actcccggcc tccctgatcagtttccctac attgatcaat 4800 ggctgagttt ggtcaggagc accccttcca tggctccactcatgcaccat tcataatttt 4860 acctccaagg tcctcctgag ccagaccgtg ttttcgcctcgaccctcagc cggttcagct 4920 cgccctgtac tgcctctctc tgaagaagag gagagtctccctcacccagt cccaccgcct 4980 taaaaccagc ctactccctt agggtcatcc catgtctcctcggctatgtc ccctgtaggc 5040 tcatcaccca ttgcctcttg gttgcaaccg tggtgggaggaagtagcccc tctactacca 5100 ctgagagagg cacaagtccc tctgggtgat gagtgctccacccccttcct ggtttatgtc 5160 ccttctttct acttctgact tgtataattg gaaaacccataatcctccct tctctgaaaa 5220 gccccaggct ttgacctcac tgatggagtc tgtactctggacacattggc ccacctggga 5280 tgactgtcaa cagctccttt tgaccctttt cacctctgaagagagggaaa gtatccaaag 5340 agaggccaaa aagtacaacc tcacatcaac caataggccggaggaggaag ctagaggaat 5400 agtgattaga gacccaattg ggacctaatt gggacccaaatttctcaagt ggagggagaa 5460 cttttgacga tttccaccgg tatctcctcg tgggtattcagggagctgct cagaaaccta 5520 taaacttgtc taaggcgact gaagtcgtcc aggggcatgatgagtcacca ggagtgtttt 5580 tagagcacct ccaggaggct tatcggattt acaccccttttgacctggca gcccccgaaa 5640 atagccatgc tcttaatttg gcatttgtgg ctcaggcagccccagatagt aaaaggaaac 5700 tccaaaaact agagggattt tgctggaatg aataccagtcagcttttaga gatagcctaa 5760 aaggtttttg acagtcaaga ggttgaaaaa caaaaacaagcagctcaggc agctgaaaaa 5820 agccactgat aaagcatcct ggagtatcag agtttactgttagatcagcc tcatttgact 5880 tcccctccca catggtgttt aaatccagct acactacttcctgactcaaa ctccactatt 5940 cctgttcatg actgtcagga actgttggaa actactgaaactggccgacc tgatcttcaa 6000 aatgtgcccc taggaaaggt ggatgccacc gtgttcacagacagtagcag cttcctcgag 6060 aagggactac gaaaggccgg tgcagctgtt accatggagacagatgtgtt gtgggctcag 6120 gctttaccag caaacacctc agcacaaaag gctgaattgatcgccctcac tcaggctctc 6180 cgatggggta aggatattaa cgttaacact gacagcaggtacgcctttgc tactgtgcat 6240 gtacgtggag ccatctacca ggagcgtggg ctactcacctcagcaggtgg ctgtaatcca 6300 ctgtaaagga catcaaaagg aaaacacggc tgttgcccgtggtaaccaga aagctgattc 6360 agcagctcaa gatgcagtgt gactttcagt cacgcctctaaacttgctgc ccacagtctc 6420 ctttccacag ccagatctgc ctgacaatcc cgcatactcaacagaagaag aaaactggcc 6480 tcagaactca gagccaataa aaatcaggaa ggttggtggattcttcctga ctctagaatc 6540 ttcatacccc gaactcttgg gaaaacttta atcagtcacctacagtctac cacccattta 6600 ggaggagcaa agctacctca gctcctccgg agccgttttaagatccccca tcttcaaagc 6660 ctaacagatc aagcagctct ccggtgcaca acctgcgcccaggtaaatgc caaaaaaggt 6720 cctaaaccca gcccaggcca ccgtctccaa gaaaactcaccaggagaaaa gtgggaaatt 6780 gactttacag aagtaaaacc acaccgggct gggtacaaataccttctagt actggtagac 6840 accttctctg gatggactga agcatttgct accaaaaacgaaactgtcaa tatggtagtt 6900 aagtttttac tcaatgaaat catccctcga cgtgggctgcctgttgccat agggtctgat 6960 aatggaccgg ccttcgcctt gtctatagtt tagtcagtcagtaaggcgtt aaacattcaa 7020 tggaagctcc attgtgccta tcgaccccag agctctgggcaagtagaacg catgaactgc 7080 accctaaaaa acactcttac aaaattaatc ttagaaaccggtgtaaattg tgtaagtctc 7140 cttcctttag ccctacttag agtaaggtgc accccttactgggctgggtt cttacctttt 7200 gaaatcatgt atgggagggc gctgcctatc ttgcctaagctaagagatgc ccaattggca 7260 aaaatatcac aaactaattt attacagtac ctacagtctccccaacaggt acaagatatc 7320 atcctgccac ttgttcgagg aacccatccc aatccaattcctgaacagac agggccctgc 7380 cattcattcc cgccaggtga cctgttgttt gttaaaaagttccagagaga aggactccct 7440 cctgcttgga agagacctca caccgtcatc acgatgccaacggctctgaa ggtggatggc 7500 attcctgcgt ggattcatca ctcccgcatc aaaaaggccaacggagccca actagaaaca 7560 tgggtcccca gggctgggtc aggcccctta aaactgcacctaagttgggt gaagccatta 7620 gattaattct ttttcttaat tttgtaaaac aatgcatagcttctgtcaaa cttatgtatc 7680 ttaagactca atataacccc cttgttataa ctgaggaatcaatgatttga ttccccaaaa 7740 acacaagtgg ggaatgtagt gtccaacctg gtttttactaaccctgtttt tagactctcc 7800 ctttccttta atcactcagc cttgtttcca cctgaattgactctccctta gctaagagcg 7860 ccagatggac tccatcttgg ctctttcact ggcagccgcttcctcaagga cttaacttgt 7920 gcaagctgac tcccagcaca tccaagaatg caattaactgataagatact gtggcaagct 7980 atatccgcag ttcccaggaa ttcgtccaat tgattacacccaaaagcccc gcgtctatca 8040 ccttgtaata atcttaaagc ccctgcacct ggaactattaacgttcctgt aaccatttat 8100 ccttttaact tttttgccta ctttatttct gtaaaattgttttaactaga ccccccctct 8160 cctttctaaa ccaaagtata aaagcaaatc tagccccttcttcaggccga gagaatttcg 8220 agcgttagcc gtctcttggc caccagctaa ataaacggattcttcatgtg tctcaaagtg 8280 tggcgttttc tctaactcgc tcaggtacga ccgtggtagtattttcccca acgtcttatt 8340 tttagggcac gtatgtagag taacttttat gaaagaaaccagttaaggag gttttgggat 8400 ttcctttatc aactgtaata ctggttttga ttatttatttatttatttat tttttttgag 8460 aaggagtttc actcttgttg cccaggctgg agtgcaatggtgcgatcttg gctcactgca 8520 acttccgcct cccaggttca agcgattctc ctgcctcagcctcgagagta gctgggatta 8580 taggcatgcg ccaccacacc cagctaattt tgtatttttagtaaagatgg ggtttcttca 8640 tgttggtcaa gctggtctgg aactccccgc ctcgggtgatctgcccgcct cggcctccga 8700 aagtgctggg attacaggtg tgatccacca cacccagccgatttatatgt atataaatca 8760 cattcctcta accaaaatgt agtgtttcct tccatcttgaatataggctg tagaccccgt 8820 gggtatggga cattgttaac agtgagacca cagcagtttttatgtcatct gacagcatct 8880 ccaaatagcc ttcatggttg tcactgcttc ccaagacaattccaaataac acttcccagt 8940 gatgacttgc tacttgctat tgttacttaa tgtgttaaggtggctgttac agacactatt 9000 agtatgtcag gaattacacc aaaatttagt ggctcaaacaatcattttat tatgtatgtg 9060 gattctcatg gtcaggtcag gatttcagac agggcacaagggtagcccac ttgtctctgt 9120 ctatgatgtc tggcctcagc acaggagact caacagctggggtctgggac catttggagg 9180 cttgttccct cacatctgat acctggcttg ggatgttggaagagggggtg agctgagact 9240 gagtgcctat atgtagtgtt tccatatggc cttgacttccttacagcctg gcagcctcag 9300 ggtagtcaga attcttagga ggcacagggc tccagggcagatgctgaggg gtcttttatg 9360 aggtagcaca gcaaatccac ccaggatc 9388 <210> SEQID NO 142 <211> LENGTH: 419 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 142 tgtaagtcga gcagtgtgat ggaaggaatg gtctttggagagagcatatc catctcctcc 60 tcactgcctc ctaatgtcat gaggtacact gagcagaattaaacagggta gtcttaacca 120 cactattttt agctaccttg tcaagctaat ggttaaagaacacttttggt ttacacttgt 180 tgggtcatag aagttgcttt ccgccatcac gcaataagtttgtgtgtaat cagaaggagt 240 taccttatgg tttcagtgtc attctttagt taacttgggagctgtgtaat ttaggctttg 300 cgtattattt cacttctgtt ctccacttat gaagtgattgtgtgttcgcg tgtgtgtgcg 360 tgcgcatgtg cttccggcag ttaacataag caaatacccaacatcacact gctcgactt 419 <210> SEQ ID NO 143 <211> LENGTH: 402 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 143 tgtaagtcgagcagtgtgat gtccactgca gtgtgttgct gggaacagtt aatgagcaaa 60 ttgtatacaatggctagtac attgaccggg atttgttgaa gctggtgagt gttatgactt 120 agcctgttagactagtctat gcacatggct ctggtcaact accgctctct catttctcca 180 gataaatcccccatgcttta tattctcttc caaacatact atcctcatca ccacatagtt 240 cctttgttaatgctttgttc tagactttcc cttttctgtt ttcttattca aacctatatc 300 tctttgcatagattgtaaat tcaaatgccc tcagggtgca ggcagttcat gtaagggagg 360 gaggctagccagtgagatct gcatcacact gctcgactta ca 402 <210> SEQ ID NO 144 <211>LENGTH: 224 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:144 tcgggtgatg cctcctcagg ccaagaagat aaagcttcag acccctaaca catttccaaa 60aaggaagaaa ggagaaaaaa gggcatcatc cccgttccga agggtcaggg aggaggaaat 120tgaggtggat tcacgagttg cggacaactc ctttgatgcc aagcgaggtg cagccggaga 180ctggggagag cgagccaatc aggttttgaa gttcctctca gtgc 224 <210> SEQ ID NO 145<211> LENGTH: 111 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 145 agccatttac cacccatcca caaaaaaaaa aaaaaaaaag aaaaatatcaaggaataaaa 60 atagactttg aacaaaaagg aacatttgct ggcctgagga ggcatcaccc g111 <210> SEQ ID NO 146 <211> LENGTH: 585 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 146 tagcatgttg agcccagacacttgtagaga gaggaggaca gttagaagaa gaagaaaagt 60 ttttaaatgc tgaaagttactataagaaag ctttggcttt ggatgagact tttaaagatg 120 cagaggatgc tttgcagaaacttcataaat atatgcaggt gattccttat ttcctcctag 180 aaatttagtg atatttgaaataatgcccaa acttaatttt ctcctgagga aaactattct 240 acattactta agtaaggcattatgaaaagt ttctttttag gtatagtttt tcctaattgg 300 gtttgacatt gcttcatagtgcctctgttt ttgtccataa tcgaaagtaa agatagctgt 360 gagaaaacta ttacctaaatttggtatgtt gttttgagaa atgtccttat agggagctca 420 cctggtggtt tttaaattattgttgctact ataattgagc taattataaa aacctttttg 480 agacatattt taaattgtcttttcctgtaa tactgatgat gatgttttct catgcatttt 540 cttctgaatt gggaccattgctgctgtgtc tgggctcaca tgcta 585 <210> SEQ ID NO 147 <211> LENGTH: 579<212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 383, 453, 465, 501 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 147 tagcatgttg agcccagacactgggcagcg ggggtggcca cggcagctcc tgccgagccc 60 aagcgtgttt gtctgtgaaggaccctgacg tcacctgcca ggctagggag gggtcaatgt 120 ggagtgaatg ttcaccgactttcgcaggag tgtgcagaag ccaggtgcaa cttggtttgc 180 ttgtgttcat cacccctcaagatatgcaca ctgctttcca aataaagcat caactgtcat 240 ctccagatgg ggaagactttttctccaacc agcaggcagg tccccatcca ctcagacacc 300 agcacgtcca ccttctcgggcagcaccacg tcctccacct tctgctggta cacggtgatg 360 atgtcagcaa agccgttctgcangaccagc tgccccgtgt gctgtgccat ctcactggcc 420 tccaccgcgt acaccgctctaggccgcgca tantgtgcac agaanaaatg atgatccagt 480 cccacagccc acgtccaagangactttatc cgtcagggat tctttattct gcaggatgac 540 ctgtggtatt aattgttcgtgtctgggctc aacatgcta 579 <210> SEQ ID NO 148 <211> LENGTH: 249 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 148 tgacaccttgtccagcatct gcaagccagg aagagagtcc tcaccaagat ccccaccccg 60 ttggcaccaggatcttggac ttccaatctc cagaactgtg agaaataagt atttgtcgct 120 aaataaatctttgtggtttc agatatttag ctatagcaga tcaggctgac taagagaaac 180 cccataagagttacatactc attaatctcc gtctctatcc ccaggtctca gatgctggac 240 aaggtgtca 249<210> SEQ ID NO 149 <211> LENGTH: 255 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 149 tgacaccttg tccagcatct gctattttgtgactttttaa taatagccat tctgactggt 60 gtgagatggt aactcattgt gggtttggtctgcatttctc taatgatcag tgatattaag 120 ctttttttaa atatgcttgt tgaccacatgtatatcatct tttgagaagt gtctgttcat 180 atcctttgcc cactttttaa tttttttatcttgtaaattt gtttaatttc cttacagatg 240 ctggacaagg tgtca 255 <210> SEQ IDNO 150 <211> LENGTH: 318 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 150 ttacgctgca acactgtgga ggccaagctg ggatcacttcttcattctaa ctggagagga 60 gggaagttca agtccagcag agggtgggtg ggtagacagtggcactcaga aatgtcagct 120 ggacccctgt ccccgcatag gcaggacagc aaggctgtggctctccaggg ccagctgaag 180 aacaggacac tgtctccgct gccacaaagc gtcagagactcccatctttg aagcacggcc 240 ttcttggtct tcctgcactt ccctgttctg ttagagacctggttatagac aaggcttctc 300 cacagtgttg cagcgtaa 318 <210> SEQ ID NO 151<211> LENGTH: 323 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 2, 7, 10, 13, 14,23, 26, 32, 44, 54, 56, 67, 74, 75, 81, 87, 104, 105, 109, 111, 120,123, 124, 136, 137, 138, 151, 155, 162, 168, 171, 176, 184, 186, 196,215, 231, 239, 252, 265, 288, 318 <223> OTHER INFORMATION: n = A,T,C orG <400> SEQUENCE: 151 tnacgcngcn acnntgtaga ganggnaagg cnttccccacattncccctt catnanagaa 60 ttattcnacc aagnntgacc natgccnttt atgacttacatgcnnactnc ntaatctgtn 120 tcnngcctta aaagcnnntc cactacatgc ntcancactgtntgtgtnac ntcatnaact 180 gtcngnaata ggggcncata actacagaaa tgcanttcatactgcttcca ntgccatcng 240 cgtgtggcct tncctactct tcttntattc caagtagcatctctggantg cttccccact 300 ctccacattg ttgcagcnat aat 323 <210> SEQ ID NO152 <211> LENGTH: 311 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 152 tcaagattcc ataggctgac cagtccaagg agagttgaaa tcatgaaggagagtctatct 60 ggagagagct gtagttttga gggttgcaaa gacttaggat ggagttggtgggtgtggtta 120 gtctctaagg ttgattttgt tcataaattt catgccctga atgccttgcttgcctcaccc 180 tggtccaagc cttagtgaac acctaaaagt ctctgtcttc ttgctctccaaacttctcct 240 gaggatttcc tcagattgtc tacattcaga tcgaagccag ttggcaaacaagatgcagtc 300 cagagggtca g 311 <210> SEQ ID NO 153 <211> LENGTH: 332<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 153caagattcca taggctgacc aggaggctat tcaagatctc tggcagttga ggaagtctct 60ttaagaaaat agtttaaaca atttgttaaa atttttctgt cttacttcat ttctgtagca 120gttgatatct ggctgtcctt tttataatgc agagtgggaa ctttccctac catgtttgat 180aaatgttgtc caggctccat tgccaataat gtgttgtcca aaatgcctgt ttagttttta 240aagacggaac tccacccttt gcttggtctt aagtatgtat ggaatgttat gataggacat 300agtagtagcg gtggtcagcc tatggaatct tg 332 <210> SEQ ID NO 154 <211>LENGTH: 345 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 154, 224, 297, 330 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 154 tcaagattccataggctgac ctggacagag atctcctggg tctggcccag gacagcaggc 60 tcaagctcagtggagaaggt ttccatgacc ctcagattcc cccaaacctt ggattgggtg 120 acattgcatctcctcagaga gggaggagat gtangtctgg gcttccacag ggacctggta 180 ttttaggatcagggtaccgc tggcctgagg cttggatcat tcanagcctg ggggtggaat 240 ggctggcagcctgtggcccc attgaaatag gctctggggc actccctctg ttcctanttg 300 aacttgggtaaggaacagga atgtggtcan cctatggaat cttga 345 <210> SEQ ID NO 155 <211>LENGTH: 295 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 46, 199, 252, 266 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 155 gacgcttggccacttgacac attaaacagt tttgcataat cactancatg tatttctagt 60 ttgctgtctgctgtgatgcc ctgccctgat tctctggcgt taatgatggc aagcataatc 120 aaacgctgttctgttaattc caagttataa ctggcattga ttaaagcatt atctttcaca 180 actaaactgttcttcatana acagcccata ttattatcaa attaagagac aatgtattcc 240 aatatcctttanggccaata tatttnatgt cccttaatta agagctactg tccgt 295 <210> SEQ ID NO156 <211> LENGTH: 406 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 172, 178, 332,338, 342, 381, 400, 402 <223> OTHER INFORMATION: n = A,T,C or G <400>SEQUENCE: 156 gacgcttggc cacttgacac tgcagtggga aaaccagcat gagccgctgcccccaaggaa 60 cctcgaagcc caggcagagg accagccatc ccagcctgca ggtaaagtgtgtcacctgtc 120 aggtgggctt ggggtgagtg ggtgggggaa gtgtgtgtgc aaagggggtgtnaatgtnta 180 tgcgtgtgag catgagtgat ggctagtgtg actgcatgtc agggagtgtgaacaagcgtg 240 cgggggtgtg tgtgcaagtg cgtatgcata tgagaatatg tgtctgtggatgagtgcatt 300 tgaaagtctg tgtgtgtgcg tgtggtcatg anggtaantt antgactgcgcaggatgtgt 360 gagtgtgcat ggaacactca ntgtgtgtgt caagtggccn ancgtc 406<210> SEQ ID NO 157 <211> LENGTH: 208 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:115, 119, 182, 187 <223> OTHER INFORMATION: n = A,T,C or G <400>SEQUENCE: 157 tgacgcttgg ccacttgaca cactaaaggg tgttactcat cactttcttctctcctcggt 60 ggcatgtgag tgcatctatt cacttggcac tcatttgttt ggcagtgactgtaanccana 120 tctgatgcat acaccagctt gtaaattgaa taaatgtctc taatactatgtgctcacaat 180 anggtanggg tgaggagaag gggagaga 208 <210> SEQ ID NO 158<211> LENGTH: 547 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 235 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 158 cttcaacctc cttcaacctccttcaacctc ctggattcaa acaatcatcc cacctcagac 60 tccttagtag ctgagactacagactcacgc cactacatct ggctaaattt ttgtagagat 120 agggtttcat catgttgccctggctggtct caaactcctg acctcaagca atgtgcccac 180 ctcagcctcc caaagtgctgggattacagg cataagccac catgcccagt ccatntttaa 240 tctttcctac cacattcttaccacactttc ttttatgttt agatacataa atgcttacca 300 ttatgataca attgcccacagtattaagac agtaacatgc tgcacaggtt tgtagcctag 360 gaacagtagg caataccacatagcttaggt gtgtggtaga ctataccatc taggtttgtg 420 taagttacac tttatgctgtttacacaatg acaaaaccat ctaatgatgc atttctcaga 480 atgtatcctt gtcagtaagctatgatgtac agggaacact gcccaaggac acagatattg 540 tacctgt 547 <210> SEQ IDNO 159 <211> LENGTH: 203 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 159 gctcctcttg ccttaccaac tcacccagta tgtcagcaattttatcrgct ttacctacga 60 aacagcctgt atccaaacac ttaacacact cacctgaaaagttcaggcaa caatcgcctt 120 ctcatgggtc tctctgctcc agttctgaac ctttctcttttcctagaaca tgcatttarg 180 tcgatagaag ttcctctcag tgc 203 <210> SEQ ID NO160 <211> LENGTH: 402 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 160 tgtaagtcga gcagtgtgat gggtggaaca gggttgtaag cagtaattgcaaactgtatt 60 taaacaataa taataatatt tagcatttat agagcacttt atatcttcaaagtacttgca 120 aacattayct aattaaatac cctctctgat tataatctgg atacaaatgcacttaaactc 180 aggacagggt catgagaraa gtatgcattt gaaagttggt gctagctatgctttaaaaac 240 ctatacaatg atgggraagt tagagttcag attctgttgg actgtttttgtgcatttcag 300 ttcagcctga tggcagaatt agatcatatc tgcactcgat gactytgcttgataacttat 360 cactgaaatc tgagtgttga tcatcacact gctcgactta ca 402 <210>SEQ ID NO 161 <211> LENGTH: 193 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 161 agcatgttga gcccagacac tgaccaggag aaaaaccaaccaatagaaac acgcccagac 60 actgaccagg agaaaaacca accaataaaa acaggcccggacataagaca aataataaaa 120 ttagcggaca aggacatgaa aacagctatt gtaagagcggatatagtggt gtgtgtctgg 180 gctcaacatg cta 193 <210> SEQ ID NO 162 <211>LENGTH: 147 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:162 tgttgagccc agacactgac caggagaaaa accaaccaat aaaaacaggc ccggacataa 60gacaaataat aaaattagcg gacaaggaca tgaaaacagc tattgtaaga gcggatatag 120tggtgtgtgt ctgggctcaa catgcta 147 <210> SEQ ID NO 163 <211> LENGTH: 294<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 163tagcatgttg agcccagaca caaatctttc cttaagcaat aaatcatttc tgcatatgtt 60tttaaaacca cagctaagcc atgattattc aaaaggacta ttgtattggg tattttgatt 120tgggttctta tctccctcac attatcttca tttctatcat tgacctctta tcccagagac 180tctcaaactt ttatgttata caaatcacat tctgtctcaa aaaatatctc acccacttct 240cttctgtttc tgcgtgtgta tgtgtgtgtg tgtgtgtctg ggctcaacat gcta 294 <210>SEQ ID NO 164 <211> LENGTH: 412 <212> TYPE: DNA <213> ORGANISM: Homosapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 292<223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 164 cgggattggctttgagctgc agatgctgcc tgtgaccgca cccggcgtgg aacagaaagc 60 cacctggctgcaagtgcgcc agagccgccc tgactacgtg ctgctgtggg gctggggcgt 120 gatgaactccaccgccctga aggaagccca ggccaccgga tacccccgcg acaagatgta 180 cggcgtgtggtgggccggtg cggagcccga tgtgcgtgac gtgggcgaag gcgccaaggg 240 ctacaacgcgctggctctga acggctacgg cacgcagtcc aaggtgatcc angacatcct 300 gaaacacgtgcacgacaagg gccagggcac ggggcccaaa gacgaagtgg gctcggtgct 360 gtacacccgcggcgtgatca tccagatgct ggacaaggtg tcaatcacta at 412 <210> SEQ ID NO 165<211> LENGTH: 361 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 165 ttgacacctt gtccagcatc tgcatctgat gagagcctca gatggctaccactaatggca 60 gaaggcaaag gagaacaggc attgtatggc aagaaaggaa gaaagagagaggggagaaag 120 gtgctaggtt cttttcaaca accagttctt gatggaactg agagtaagagctcaaggcca 180 ggtgtggtga ctccaaccag taatcccaac attttaggag gctgaggcaggcagatgtct 240 tgaccccatg agtttgtgac cagcctgaac aacatcatga gactccatctctacaataat 300 tacaaaaatt aatcaggcat tgtggtatgc cctgtagtcc cagatgctggacaaggtgtc 360 a 361 <210> SEQ ID NO 166 <211> LENGTH: 427 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 166 twgactgactcatgtcccct acacccaact atcttctcca ggtggccagg catgatagaa 60 tctgatcctgacttagggga atattttctt tttacttccc atcttgattc cctgccggtg 120 agtttcctggttcagggtaa gaaaggagct caggccaaag taatgaacaa atccatcctc 180 acagacgtacagaataagag aacwtggacw tagccagcag aacmcaaktg aaamcagaac 240 mcttamctaggatracaamc mcrraratar ktgcycmcmc wtataataga aaccaaactt 300 gtatctaattaaatatttat ccacygtcag ggcattagtg gttttgataa atacgctttg 360 gctaggattcctgaggttag aatggaaraa caattgcamc gagggtaggg gacatgagtc 420 aktctaa 427<210> SEQ ID NO 167 <211> LENGTH: 500 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:288, 303, 318, 326 <223> OTHER INFORMATION: n = A,T,C or G <400>SEQUENCE: 167 aacgtcgcat gctcccggcc gccatggccg cgggatagac tgactcatgtcccctaagat 60 agaggagaca cctgctaggt gtaaggagaa gatggttagg tctacggaggctccagggtg 120 ggagtagttc cctgctaagg gagggtagac tgttcaacct gttcctgctccggcctccac 180 tatagcagat gcgagcagga gtaggagaga gggaggtaag agtcagaagcttatgttgtt 240 tatgcgggga aacgccrtat cgggggcagc cragttatta ggggacantrtagwyartcw 300 agntagcatc caaagcgngg gagttntccc atatggttgg acctgcaggcggccgcatta 360 gtgattagca tgtgagcccc agacacgcat agcaacaagg acctaaactcagatcctgtg 420 ctgattactt aacatgaatt attgtattta tttaacaact ttgagttatgaggcatatta 480 ttaggtccat attacctgga 500 <210> SEQ ID NO 168 <211>LENGTH: 358 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:168 ttcatcgctc ggtgactcaa gcctgtaatc ccagaacttt gggaggccga ggggagcaga 60tcacctgagg ttgggagttt gagaccagcc tggccaacat ggtgacaacc cgtctctgct 120aaaaatacaa aaattagcca agcatggtgg catgcacttg taatcccagc tactcgggag 180gctgaggcag gagaatcact tgaggccagg aggcagaggt tgcagtgagg cagaggttga 240gatcatgcca ctgcactcca gcctgggcaa cagagtaaga ctccatctca aaaaaaaaaa 300aaaaaaagaa tgatcagagc cacaaataca gaaaaccttg agtcaccgag cgatgaaa 358<210> SEQ ID NO 169 <211> LENGTH: 1265 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 169 ttctgtccac accaatctta gagctctgaaagaatttgtc tttaaatatc ttttaatagt 60 aacatgtatt ttatggacca aattgacattttcgactatt ttttcccaaa aaaagtcagg 120 tgaatttcag cacactgagt tgggaatttcttatcccaga agwcggcacg agcaatttca 180 tatttattta agattgattc catactccgttttcaaggag aatccctgca gtctccttaa 240 aggtagaaca aatactttct atttttttttcaccattgtg ggattggact ttaagaggtg 300 actctaaaaa aacagagaac aaatatgtctcagttgtatt aagcacggac ccatattatc 360 atattcactt aaaaaaatga tttcctgtgcaccttttggc aacttctctt ttcaatgtag 420 ggaaaaactt agtcaccctg aaaacccacaaaataaataa aacttgtaga tgtgggcaga 480 argtttgggg gtggacattg tatgtgtttaaattaaaccc tgtatcactg agaagctgtt 540 gtatgggtca gagaaaatga atgcttagaagctgttcaca tcttcaagag cagaagcaaa 600 ccacatgtct cagctatatt attatttattttttatgcat aaagtgaatc atttcttctg 660 tattaatttc caaagggttt taccctctatttaaatgctt tgaaaaacag tgcattgaca 720 atgggttgat atttttcttt aaaagaaaaatataattatg aaagccaaga taatctgaag 780 cctgttttat tttaaaactt tttatgttctgtggttgatg ttgtttgttt gtttgtttct 840 attttgttgg ttttttactt tgttttttgttttgttttgt tttggtttdg catactacat 900 gcagtttctt taaccaatgt ctgtttggctaatgtaatta aagttgttaa tttatatgag 960 tgcatttcaa ctatgtcaat ggtttcttaatatttattgt gtagaagtac tggtaatttt 1020 tttatttaca atatgtttaa agagataacagtttgatatg ttttcatgtg tttatagcag 1080 aagttattta tttctatggc attccagcggatattttggt gtttgcgagg catgcagtca 1140 atattttgta cagttagtgg acagtattcagcaacgcctg atagcttctt tggccttatg 1200 ttaaataaaa agacctgttt gggatgtaaaaaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 1260 aaaaa 1265 <210> SEQ ID NO 170<211> LENGTH: 383 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 170 tgtaagtcga gcagtgtgat gacgatattc ttcttattaa tgtggtaattgaacaaatga 60 tctgtgatac tgatcctgag ctaggaggcg ctgttcagtt aatgggacttcttcgtactc 120 taattgatcc agagaacatg ctggctacaa ctaataaaac cgaaaaaagtgaatttctaa 180 attttttcta caaccattgt atgcatgttc tcacagcacc acttttgaccaatacttcag 240 aagacaaatg tgaaaaggat aatatagttg gatcaaacaa aaacaacacaatttgtcccg 300 ataattatca aacagcacag ctacttgcct taattttaga gttactcacattttgtgtgg 360 aacatcacac tgctcgactt aca 383 <210> SEQ ID NO 171 <211>LENGTH: 383 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:171 tgggcacctt caatatcgca agttaaaaat aatgttgagt ttattatact tttgacctgt 60ttagctcaac agggtgaagg catgtaaaga atgtggactt ctgaggaatt ttcttttaaa 120aagaacataa tgaagtaaca ttttaattac tcaaggacta cttttggttg aagtttataa 180tctagatacc tctacttttt gtttttgctg ttcgacagtt cacaaagacc ttcagcaatt 240tacagggtaa aatcgttgaa gtagtggagg tgaaactgaa atttaaaatt attctgtaaa 300tactataggg aaagaggctg agcttagaat cttttggttg ttcatgtgtt ctgtgctctt 360atcatcacac tgctcgactt aca 383 <210> SEQ ID NO 172 <211> LENGTH: 699<212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 641 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 172 tcgggtgatg cctcctcagg cttgtcgttagtgtacacag agctgctcat gaagcgacag 60 cggctgcccc tggcacttca gaacctcttcctctacactt ttggtgcgct tctgaatcta 120 ggtctgcatg ctggcggcgg ctctggcccaggcctcctgg aaagtttctc aggatgggca 180 gcactcgtgg tgctgagcca ggcactaaatggactgctca tgtctgctgt catggagcat 240 ggcagcagca tcacacgcct ctttgtggtgtcctgctcgc tggtggtcaa cgccgtgctc 300 tcagcagtcc tgctacggct gcagctcacagccgccttct tcctggccac attgctcatt 360 ggcctggcca tgcgcctgta ctatggcagccgctagtccc tgacaacttc caccctgatt 420 ccggaccctg tagattgggc gccaccaccagatccccctc ccaggccttc ctccctctcc 480 catcagcggc cctgtaacaa gtgccttgtgagaaaagctg gagaagtgag ggcagccagg 540 ttattctctg gaggttggtg gatgaaggggtacccctagg agatgtgaag tgtgggtttg 600 gttaaggaaa tgcttaccat cccccacccccaaccaagtt nttccagact aaagaattaa 660 ggtaacatca atacctaggc ctgaggaggcatcacccga 699 <210> SEQ ID NO 173 <211> LENGTH: 701 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 173 tcgggtgatg cctcctcaggccagatcaaa cttggggttg aaaactgtgc aaagaaatca 60 atgtcggaga aagaattttgcaaaagaaaa atgcctaatc agtactaatt taataggtca 120 cattagcagt ggaagaagaaatgttgatat tttatgtcag ctattttata atcaccagag 180 tgcttagctt catgtaagccatctcgtatt cattagaaat aagaacaatt ttattcgtcg 240 gaaagaactt ttcaatttatagcatcttaa ttgctcagga ttttaaattt tgataaagaa 300 agctccactt ttggcaggagtagggggcag ggagagagga ggctccatcc acaaggacag 360 agacaccagg gccagtagggtagctggtgg ctggatcagt cacaacggac tgacttatgc 420 catgagaaga aacaacctccaaatctcagt tgcttaatac aacacaagct catttcttgc 480 tcacgttaca tgtcctatgtagatcaacag caggtgactc agggacccag gctccatctc 540 catatgagct tccatagtcaccaggacacg ggctctgaaa gtgtcctcca tgcagggaca 600 catgcctctt cctttcattgggcagagcaa gtcacttatg gccagaagtc acactgcagg 660 gcagtgccat cctgctgtatgcctgaggag gcatcacccg a 701 <210> SEQ ID NO 174 <211> LENGTH: 700 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 19 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 174 tcgggtgatg cctcctcang cccctaaatc agagtccagggtcagagcca caggagacag 60 ggaaagacat agattttaac cggccccctt caggagattctgaggctcag ttcactttgt 120 tgcagtttga acagaggcag caaggctagt ggttaggggcacggtctcta aagctgcact 180 gcctggatct gcctcccagc tctgccagga accagctgcgtggccttgag ctgctgacac 240 gcagaaagcc ccctgtggac ccagtctcct cgtctgtaagatgaggacag gactctagga 300 accctttccc ttggtttggc ctcactttca caggctcccatcttgaactc tatctactct 360 tttcctgaaa ccttgtaaaa gaaaaaagtg ctagcctgggcaacatggca aaaccctgtc 420 tctacaaaaa atacaaaaat tagttgggtg tggtggcatgtgcctgtagt cccagccact 480 tgggaggtgc tgaggtggga ggatcacttg agcccgggaggtggaggttg cagtgagcca 540 agatcatgcc actgcactcc agcctgagta atagagtaagactctgtctc aaaaacaaca 600 acaacaacag tgagtgtgcc tctgtttccg ggttggatggggcaccacat ttatgcatct 660 ctcagatttg gacgctgcag cctgaggagg catcacccga700 <210> SEQ ID NO 175 <211> LENGTH: 484 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 30 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 175tatagggcga attgggcccg agttgcatgn tcccggccgc catggccgcg ggattcgggt 60gatgcctcct caggcttgtc tgccacaagc tacttctctg agctcagaaa gtgccccttg 120atgagggaaa atgtcctact gcactgcgaa tttctcagtt ccattttacc tcccagtcct 180ccttctaaac cagttaataa attcattcca caagtattta ctgattacct gcttgtgcca 240gggactattc tcaggctgaa gaaggtggga ggggagggcg gaacctgagg agccacctga 300gccagcttta tatttcaacc atggctggcc catctgagag catctcccca ctctcgccaa 360cctatcgggg catagcccag ggatgccccc aggcggccca ggttagatgc gtccctttgg 420cttgtcagtg atgacataca ccttagctgc ttagctggtg ctggcctgag gaggcatcac 480ccga 484 <210> SEQ ID NO 176 <211> LENGTH: 432 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 176 tcgggtgatg cctcctcagggctcaaggga tgagaagtga cttctttctg gagggaccgt 60 tcatgccacc caggatgaaaatggataggg acccacttgg aggacttgct gatatgtttg 120 gacaaatgcc aggtagcggaattggtactg gtccaggagt tatccaggat agattttcac 180 ccaccatggg acgtcatcgttcaaatcaac tcttcaatgg ccatggggga cacatcatgc 240 ctcccacaca atcgcagtttggagagatgg gaggcaagtt tatgaaaagc caggggctaa 300 gccagctcta ccataaccagagtcagggac tcttatccca gctgcaagga cagtcgaagg 360 atatgccacc tcggttttctaagaaaggac agcttaatgc agatgagatt agcctgagga 420 ggcatcaccc ga 432 <210>SEQ ID NO 177 <211> LENGTH: 788 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 177 tagcatgttg agcccagaca cagtagcatt tgtgccaatttctggttgga atggtgacaa 60 catgctggag ccaagtgcta acatgccttg gttcaagggatggaaagtca cccgtaagga 120 tggcaatgcc agtggaacca cgctgcttga ggctctggactgcatcctac caccaactcg 180 cccaactgac aagcccttgc gcctgcctct ccaggatgtctacaaaattg gtggtattgg 240 tactgttcct gttggccgag tggagactgg tgttctcaaacccggtatgg tggtcacctt 300 tgctccagtc aacgttacaa cggaagtaaa atctgtcgaaatgcaccatg aagctttgag 360 tgaagctctt cctggggaca atgtgggctt caatgtcaagaatgtgtctg tcaaggatgt 420 tcgtcgtggc aacgttgctg gtgacagcaa aaatgacccaccaatggaag cagctggctt 480 cactgctcag gtgattatcc tgaaccatcc aggccaaataagtgccggct atgcccctgt 540 attggattgc cacacggctc acattgcatg caagtttgctgagctgaagg aaaagattga 600 tcgccgttct ggtaaaaagc tggaagatgg ccctaaattcttgaagtctg gtgatgctgc 660 cattgttgat atggttcctg gcaagcccat gtgtgttgagagcttctcag actatccacc 720 tttgggtcgc tttgctgttc gtgatatgag acagacagttgcggtgggtg tctgggctca 780 acatgcta 788 <210> SEQ ID NO 178 <211> LENGTH:786 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 178tagcatgttg agcccagaca cctgtgtttc tgggagctct ggcagtggcg gattcatagg 60cacttgggct gcactttgaa tgacacactt ggctttatta gattcactag tttttaaaaa 120attgttgttc gtttcttttc attaaaggtt taatcagaca gatcagacag cataattttg 180tatttaatga cagaaacgtt ggtacatttc ttcatgaatg agcttgcatt ctgaagcaag 240agcctacaaa aggcacttgt tataaatgaa agttctggct ctagaggcca gtactctgga 300gtttcagagc agccagtgat tgttccagtc agtgatgcct agttatatag aggaggagta 360cactgtgcac tcttctaggt gtaagggtat gcaactttgg atcttaaaat tctgtacaca 420tacacacttt atatatatgt atgtatgtat gaaaacatga aattagtttg tcaaatatgt 480gtgtgtttag tattttagct tagtgcaact atttccacat tatttattaa attgatctaa 540gacactttct tgttgacacc ttgaatatta atgttcaagg gtgcaatgtg tattccttta 600gattgttaaa gcttaattac tatgatttgt agtaaattaa cttttaaaat gtatttgagc 660ccttctgtag tgtcgtaggg ctcttacagg gtgggaaaga ttttaatttt ccagttgcta 720attgaacagt atggcctcat tatatatttt gatttatagg agtttgtgtc tgggctcaac 780atgcta 786 <210> SEQ ID NO 179 <211> LENGTH: 796 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 179 tagcatgttg agcccagacactggttacaa gaccagacct gcttcctcca tatgtaaaca 60 gcttttaaaa agccagtgaacctttttaat actttggcaa ccttctttca caggcaaaga 120 acacccccat ccgccccttgtttggagtgc agagtttggc tttggttctt tgccttgcct 180 ggagtatact tctaattcctgttgtcctgc acaagctgaa taccgagcta cccaccgcca 240 cccaggccag gtttccactcatttattact ttatgtttct gttccattgc tggtccacag 300 aaataagttt tcctttggaggaatgtgatt ataccccttt aatttcctcc ttttgctttt 360 ttttaatatc attggtatgtgtttggccca gaggaaactg aaattcacca tcatcttgac 420 tggcaatccc attaccatgctttttttaaa aaacgtaatt tttcttgcct tacattggca 480 gagtagccct tcctggctactggcttaatg tagtcactca gtttctaggt ggcattaggc 540 atgagacctg aagcacagactgtcttacca caaaaggtga caagatctca aaccttagcc 600 aaagggctat gtcaggtttcaatgctatct gcttctgttc ctgctcactg ttctggattt 660 tgtccttctt catccctagcaccagaattt cccagtctcc ctccctacct tcccttgttt 720 taattctaat ctatcagcaaaataactttt caaatgtttt aaccggtatc tccatgtgtc 780 tgggctcaac atgcta 796<210> SEQ ID NO 180 <211> LENGTH: 488 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 180 ggatgtgctg caaggcgatt aagttgggtaacgccagggt tttcccagtc acgacgttgt 60 aaaacgacgg ccagtgaatt gtaatacgactcactatagg gcgaattggg cccgacgtcg 120 catgctcccg gccgccatgg ccgcgggatagcatgttgag cccagacacc tgcaggtcat 180 ttggagagat ttttcacgtt accagcttgatggtcttttt caggaggaga gacactgagc 240 actcccaagg tgaggttgaa gatttcctctagatagccgg ataagaagac taggagggat 300 gcctagaaaa tgattagcat gcaaatttctacctgccatt tcagaactgt gtgtcagccc 360 acattcagct gcttcttgtg aactgaaaagagagaggtat tgagactttt ctgatggccg 420 ctctaacatt gtaacacagt aatctgtgtgtgtgtgggtg tgtgtgtgtg tctgggctca 480 acatgcta 488 <210> SEQ ID NO 181<211> LENGTH: 317 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 181 tagcatgttg agcccagaca cggcgacggt acctgatgag tggggtgatggcacctgtga 60 aaaggaggaa cgtcatcccc catgatattg gggacccaga tgatgaaccatggctccgcg 120 tcaatgcata tttaatccat gatactgctg attggaagga cctgaacctgaagtttgtgc 180 tgcaggttta tcgggactat tacctcacgg gtgatcaaaa cttcctgaaggacatgtggc 240 ctgtgtgtct agtaagggat gcacatgcag tggccagtgt gccaggggtatggttggtgt 300 ctgggctcaa catgcta 317 <210> SEQ ID NO 182 <211> LENGTH:507 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 493 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 182 tagcatgttg agcccagaca ctggctgttagccaaatcct ctctcagctg ctccctgtgg 60 tttggtgact caggattaca gaggcatcctgtttcaggga acaaaaagat tttagctgcc 120 agcagagagc accacataca ttagaatggtaaggactgcc acctccttca agaacaggag 180 tgagggtggt ggtgaatggg aatggaagcctgcattccct gatgcatttg tgctctctca 240 aatcctgtct tagtcttagg aaaggaagtaaagtttcaag gacggttccg aactgctttt 300 tgtgtctggg ctcaacatgc tatcccgcggccatggcggc cgggagcatg cgacgtcggg 360 cccaattcgc cctatagtga gtcgtattacaattcactgg ccgtcgtttt acaacgtcgt 420 gactgggaaa accctggcgt tacccaacttaatcgccttg cagcacatcc ccctttccca 480 gctggcgtaa tancgaaaag gcccgca 507<210> SEQ ID NO 183 <211> LENGTH: 227 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 183 gatttacgct gcaacactgt ggaggtagccctggagcaag gcaggcatgg atgcttctgc 60 aatccccaaa tggagcctgg tatttcagccaggaatctga gcagagcccc ctctaattgt 120 agcaatgata agttattctc tttgttcttcaaccttccaa tagccttgag cttccagggg 180 agtgtcgtta atcattacag cctggtctccacagtgttgc agcgtaa 227 <210> SEQ ID NO 184 <211> LENGTH: 225 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 184 ttacgctgcaacactgtgga gcagattaac atcagacttt tctatcaaca tgactggggt 60 tactaaaaagacaacaaatc aatggcttca aaagtctaag gaataatttc gatacttcaa 120 ctttataaaacctgacaaaa ctatcaatca agcataaaga cagatgaaga acatttccag 180 attttggccaatcagatatt ttacctccac agtgttgcag cgtaa 225 <210> SEQ ID NO 185 <211>LENGTH: 597 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:185 ggcccgacgt cgcatgctcc cggccgccat ggccgcggga ttcgttaggg tctctatcca 60ctgggaccca taggctagtc agagtattta gagttgagtt cctttctgct tcccagaatt 120tgaaagaaaa ggagtgaggt gatagagctg agagatcaga tttgcctctg aagcctgttc 180aagatgtatg tgctcagacc ccaccactgg ggcctgtggg tgaggtcctg ggcatctatt 240tgaatgaatt gctgaagggg agcactatgc caaggaaggg gaacccatcc tggcactggc 300acaggggtca ccttatccag tgctcagtgc ttctttgctg ctacctggtt ttctctcata 360tgtgaggggc aggtaagaag aagtgcccrg tgttgtgcga gttttagaac atctaccagt 420aagtggggaa gtttcacaaa gcagcagctt tgttttgtgt attttcacct tcagttagaa 480gaggaaggct gtgagatgaa tgttagttga gtggaaaaga cgggtaagct tagtggatag 540agaccctaac gaatcactag tgcggccgcc ttgcaggtcg accatatggg agagctc 597 <210>SEQ ID NO 186 <211> LENGTH: 597 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 186 ggcccgaagt tgcatgttcc cggccgccat ggccgcgggattcgttaggg tctctatcca 60 ctacctaaaa aatcccaaac atataactga actcctcacacccaattgga ccaatccatc 120 accccagagg cctacagatc ctcctttgat acataagaaaatttccccaa actacctaac 180 tatatcattt tgcaagattt gttttaccaa attttgatggcctttctgag cttgtcagtg 240 tgaaccacta ttacgaacga tcggatatta actgcccctcaccgtccagg tgtagctggc 300 aacatcaagt gcagtaaata ttcattaagt tttcacctactaaggtgctt aaacacccta 360 gggtgccatg tcggtagcag atcttttgat ttgtttttatttcccataag ggtcctgttc 420 aaggtcaatc atacatgtag tgtgagcagc tagtcactatcgcatgactt ggagggtgat 480 aatagaggcc tcctttgctg ttaaagaact cttgtcccagcctgtcaaag tggatagaga 540 ccctaacgaa tcactagtgc ggccgcctgc aggtcgaccatatgggagag ctcccaa 597 <210> SEQ ID NO 187 <211> LENGTH: 324 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 187 tcgttagggtctctatccac ttgcaggtaa aatccaatcc tgtgtatatc ttatagtctt 60 ccatatgtagtggttcaaga gactgcagtt ccagaaagac tagccgagcc catccatgtc 120 ttccacttaaccctgctttg ggttacacat cttaactttt ctgttcaagt ttctctgtgt 180 agtttatagcatgagtattg ggawaatgcc ctgaaacctg acatgagatc tgggaaacac 240 aaacttactcaataagaatt tctcccatat ttttatgatg gaaaaatttc acatgcacag 300 aggagtggatagagacccta acga 324 <210> SEQ ID NO 188 <211> LENGTH: 178 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 46 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 188 gcgcggggat tcggggtgat acctcctcat gccaaaatacaacgtntaat ttcacaactt 60 gccttccaat ttacgcattt tcaatttgct ctccccatttgttgagtcac aacaaacacc 120 attgcccaga aacatgtatt acctaacatg cacatactcttaaaactact catccctt 178 <210> SEQ ID NO 189 <211> LENGTH: 367 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 189 tgacaccttgtccagcatct gacacagtct tggctcttgg aaaatattgg ataaatgaaa 60 atgaatttctttagcaagtg gtataagctg agaatatacg tatcacatat cctcattcta 120 agacacattcagtgtccctg aaattagaat aggacttaca ataagtgtgt tcactttctc 180 aatagctgttattcaattga tggtaggcct taaaagtcaa agaaatgaga gggcatgtga 240 aaaaaagctcaacatcactg atcattagaa aacttccatt caaaccccca atgagatacc 300 atctcataccagtcagaatg gctattatta aaaagtcaaa aaataacaga tgctggacaa 360 ggtgtca 367<210> SEQ ID NO 190 <211> LENGTH: 369 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:323 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 190gacaccttgt ccagcatctg acaacgctaa cagcctgagg agatctttat ttatttattt 60agtttttact ctggctaggc agatggtggc taaaacattc atttacccat ttattcattt 120aattgttcct gcaaggccta tggatagagt attgtccagc actgctctgg aagctaggag 180catggggatg aacaagatag gctacatcct gttcccacag aacttccact ttagtctggg 240aaacagatga tatatacaaa tatataaatg aattcaggta gttttaagta cgaaaagaat 300aagaaagcag agtcatgatt tanaatgctg gaaacagggg ctattgcttg agatattgaa 360ggtgcccaa 369 <210> SEQ ID NO 191 <211> LENGTH: 369 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 191 tgacaccttg tccagcatctgcacagggaa aagaaactat tatcagagtg aacaggcaac 60 ctacagaatg ggagaaaatttttgcaatct atccatctga caaagggcta atatccagaa 120 tctacaaaga acttatacaaatttacaaga aacaaacaaa caaacaactc ctcaaaaagt 180 gggtgaagga tgtgaacagacacttctcaa aagaagacat ttatggggcc aacaaacata 240 tgaaaaaaag ctcatcatcactggtcacta gataaatgca aatcaaaacc acaatgagat 300 accatctcat tccagttagaatggcaatca ttaaaaagtc aggaaacaac agatgctgga 360 caaggtgtc 369 <210> SEQID NO 192 <211> LENGTH: 449 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 192 tgacgcttgg ccacttgaca cttcatcttt gcacagaaaaacttctttac agatttaatt 60 caagactggt ctagtgacag tcctccagac attttttcatttgttccata tacgtggaat 120 tttaaaatca tgtttcatca gtttgaaatg atttgggctgctaatcaaca caattggatc 180 gactgttcta ctaaacaaca ggaaaatgtg tatctggcagcctgtggaga aacactaaac 240 attgattttt ctttgccttt tacggacttt gttccagctacatgtaatac caagttctct 300 ttaagaggag aagatgttga tcttcatttg tttctaccagactgccaccc tagtaaatat 360 tctttattta tgctggtaaa aaattgccat ccaaataagatgattcatga tactggtatt 420 cctgctgagt gtcaagtggc caagcgtca 449 <210> SEQID NO 193 <211> LENGTH: 372 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 193 tgacgcttgg ccacttgaca ccagggatgt akcagttgaatataatcctg caattgtaca 60 tattggcaat ttcccatcaa acattctaga aagagacaaccaggattgct aggccataaa 120 agctgcaata aataactggt aattgcagta atcatttcaggccaattcaa tccagtttgg 180 ctcagaggtg cctttggctg agagaagagg tgagatataatgtgttttct tgcaacttct 240 tggaagaata actccacaat agtctgagga ctagatacaaacctatttgc cattaaagca 300 ccagagtctg ttaattccag tactgataag tgttggagattagactccag tgtgtcaagt 360 ggccaagcgt ca 372 <210> SEQ ID NO 194 <211>LENGTH: 309 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 140, 205 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 194 tgacgcttgg ccacttgacacttatgtaga atccatcgtg ggctgatgca agccctttat 60 ttaggcttag tgttgtgggcaccttcaata tcacactaga gacaaacgcc acaagatctg 120 cagaaacatt cagttctgancactcgaatg gcaggataac tttttgtgtt gtaatccttc 180 acatatacaa aaacaaactctgcantctca cgttacaaaa aaacgtactg ctgtaaaata 240 ttaagaaggg gtaaaggataccatctataa caaagtaact tacaactagt gtcaagtggc 300 caagcgtca 309 <210> SEQID NO 195 <211> LENGTH: 312 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 100, 270<223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 195 tgacgcttggccacttgaca cccaatctcg cacttcatcc tcccagcacc tgatgaagta 60 ggactgcaactatccccact tcccagatga ggggaccaan gtacacatta ggacccggat 120 gggagcacagatttgtccga tcccagactc caagcactca gcgtcactcc aggacagcgg 180 ctttcagataaggtcacaaa catgaatggc tccgacaacc ggagtcagtc cgtgctgagt 240 taaggcaatggtgacacgga tgcacgtgtn acctgtaatg gttcatcgta agtgtcaagt 300 ggccaagcgt ca312 <210> SEQ ID NO 196 <211> LENGTH: 288 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 196 tgtatcgacg tagtggtctcctcagccatg cagaactgtg actcaattaa acctctttcc 60 tttatgaatt acccaatctcgggtagtgtc tttatagtag tgtgagaatg gactaataca 120 agtacatttt acttagtaataataataaac aaatatatta catttttgtg tatttactac 180 accatatttt ttattgttattgtagtgtac accttctact tattaaaaga aataggcccg 240 aggcgggcag atcacgaggtcaggagatgg agaccactac gtcgatac 288 <210> SEQ ID NO 197 <211> LENGTH: 289<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 197ttgggcacct tcaatatcat gacaggtgat gtgataacca agaaggctac taagtgatta 60atgggtgggt aatgtataca gagtaggtac actggacaga ggggtaattc atagccaagg 120caggagaagc agaatggcaa aacatttcat cacactactc aggatagcat gcagtttaaa 180acctataagt agtttatttt tggaattttc cacttaatat tttcagactg caggtaacta 240aactgtggaa cacaagaaca tagataaggg gagaccacta cgtcgatac 289 <210> SEQ IDNO 198 <211> LENGTH: 288 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 198 gtatcgacgt agtggtctcc caagcagtgg gaagaaaacgtgaaccaatt aaaatgtatc 60 agatacccca aagaaaggcg cttgagtaaa gattccaagtgggtcacaat ctcagatctt 120 aaaattcagg ctgtcaaaga gatttgctat gaggttgctctcaatgactt caggcacagt 180 cggcaggaga ttgaagccct ggccattgtc aagatgaaggagctttgtgc catgtatggc 240 aagaaagacc ccaatgagcg ggactcctgg agaccactacgtcgatac 288 <210> SEQ ID NO 199 <211> LENGTH: 1027 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature<222> LOCATION: 17, 21, 36, 39, 40, 42, 63, 98, 116, 145, 162, 173, 865,885, 891, 916, 924, 927, 929, 934, 942, 949, 976, 983, 988, 989, 1009,1014 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 199gctttttggg aaaaacncaa ntgggggaaa gggggnttnn tngcaagggg ataaaggggg 60aancccaggg tttccccatt cagggaggtg taaaaagncg gccaggggat tgtaanagga 120ttcaataata gggggaatgg gcccngaagt tgcaaggttc cngcccgcca tgnccgcggg 180atttagtgac attacgacgs tggtaataaa gtgggsccaa waaatatttg tgatgtgatt 240tttsgaccag tgaacccatt gwacaggacc tcatttccty tgagatgrta gccataatca 300gataaaagrt tagaagtytt tctgcacgtt aacagcatca ttaaatggag tggcatcacc 360aatttcaccc tttgttagcc gataccttcc ccttgaaggc attcaattaa gtgaccaatc 420gtcatacgag aggggatggc atggggattg atgatgatat caggggtgat accttcacag 480gtgaaaggca tatcctcttg tctatactga ataccacaag tacccttttg accatgtcga 540ctagcaaatt tgtctccaat ctgtgtwatc cctaacagag cgtaccctta ttttacaaaa 600tttatatcct tcctgattga gagttaccat aacctgatcc acaatgcccg tctcgctwgt 660tctgagaaaa gtgctacagt ctctcttggt atagcgtcta ttggtgctct ccaattcatc 720ttcatttttc aggcaaggtg aactgttttg cctataataa cmtcatctcc tgatacmcga 780aacccckgga rctatcaaac catcatcatc cagcgttckt watgtymcta aatccctatt 840gcggccgcct gcaggtcaac atatnggaaa accccccacc ccttnggagc ntaccttgaa 900ttttccatat gtcccntaaa ttanctngnc ttancctggc cntaacctnt tccggtttaa 960attgtttccg cccccnttcc ccnccttnna accggaaacc ttaattttna accnggggtt 1020cctatcc 1027 <210> SEQ ID NO 200 <211> LENGTH: 207 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 200 agtgacatta cgacgctggccatcttgaat cctagggcat gaagttgccc caaagttcag 60 cacttggtta agcctgatccctctggttta tcacaaagaa taggatggga taaagaaagt 120 ggacacttaa ataagctataaattatatgg tccttgtcta gcaggagaca actgcacagg 180 tatactacca gcgtcgtaatgtcacta 207 <210> SEQ ID NO 201 <211> LENGTH: 209 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 201 tgggcacctt caatatctattaaaagcaca aatactgaag aacacaccaa gactatcaat 60 gaggttacat ctggagtcctcgatatatca ggaaaaaatg aagtgaacat tcacagagtt 120 ttacttcttt gggaactcaaatgctagaaa agaaaagggt gccctctttc tctggcttcc 180 tggtcctatc cagcgtcgtaatgtcacta 209 <210> SEQ ID NO 202 <211> LENGTH: 349 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature<222> LOCATION: 1 <223> OTHER INFORMATION: n = A,T,C or G <400>SEQUENCE: 202 ntacgctgca acactgtgga gccactggtt tttattcccg gcaggttatccagcaaacag 60 tcactgaaca caccgaagac cgtggtatgg taaccgttca cagtaatcgttccagtcgtc 120 tgcgggaccc cgacgagcgt cactgggtac agaccagatt cagccggaagagaaagcgcc 180 gcagggagag actcgaactc cactccgctg gtgagcagcc ccatgttttcaactcgaagt 240 tcaaacggca ttgggttata taccatcagc tgaacttcac acacatctccttgaacccac 300 tggaaatcta ttttcttgtt ccgctcttct ccacagtgtt gcagcgtaa 349<210> SEQ ID NO 203 <211> LENGTH: 241 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 203 tgctcctctt gccttaccaa cccaaagcccactgtgaaat atgaagtgaa tgacaaaatt 60 cagttttcaa cgcaatatag tatagtttatctgattcttt tgatctccag gacactttaa 120 acaactgcta ccaccaccac caacctagggatttaggatt ctccacagac cagaaattat 180 ttctcctttg agtttcaggc tcctctgggactcctgttca tcaatgggtg gtaaatggct 240 a 241 <210> SEQ ID NO 204 <211>LENGTH: 248 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:204 tagccattta ccacccatct gcaaaccswg acmwwcargr cywgwackya ggcgatttga 60agtactggta atgctctgat catgttagtt acataagtgt ggtcagttta caaaaattca 120cagaactaaa tactcaatgc tatgtgttca tgtctgtgtt tatgtgtgtg taatgtttca 180attaagtttt tttaaaaaaa agagatgatt tccaaataag aaagccgtgt tggtaaggca 240agaggagc 248 <210> SEQ ID NO 205 <211> LENGTH: 505 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 447 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE:205 tacgctgcaa cactgtggag ccattcatac aggtccctaa ttaaggaaca agtgattatg 60ctacctttgc acggttaggg taccgcggcc gttaaacatg tgtcactggg caggcggtgc 120ctctaatact ggtgatgcta gaggtgatgt ttttggtaaa caggcggggt aagatttgcc 180gagttccttt tacttttttt aacctttcct tatgagcatg cctgtgttgg gttgacagtg 240ggggtaataa tgacttgttg gttgattgta gatattgggc tgttaattgt cagttcagtg 300ttttaatctg acgcaggctt atgcggagga gaatgttttc atgttactta tactaacatt 360agttcttcta tagggtgata gattggtcca attgggtgtg aggagttcag ttatatgttt 420gggatttttt aggtagtggg tgttganctt gaacgctttc ttaattggtg gctgctttta 480rgcctactat gggtggtaaa tggct 505 <210> SEQ ID NO 206 <211> LENGTH: 179<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 206tagactgact catgtcccct accaaagccc atgtaaggag ctgagttctt aaagactgaa 60gacagactat tctctggaga aaaataaaat ggaaattgta ctttaaaaaa aaaaaaaatc 120ggccgggcat ggtagcacac acctgtaatc ccagctacta ggggacatga gtcagtcta 179<210> SEQ ID NO 207 <211> LENGTH: 176 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 207 agactgactc atgtccccta ccccaccttctgctgtgctg ccgtgttcct aacaggtcac 60 agactggtac tggtcagtgg cctgggggttggggacctct attatatggg atacaaattt 120 aggagttgga attgacacga tttagtgactgatgggatat gggtggtaaa tggcta 176 <210> SEQ ID NO 208 <211> LENGTH: 196<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 208agactgactc atgtccccta tttaacaggg tctctagtgc tgtgaaaaaa aaaaatgctg 60aacattgcat ataacttata ttgtaagaaa tactgtacaa tgactttatt gcatctgggt 120agctgtaagg catgaaggat gccaagaagt ttaaggaata tgggtggtaa atggctaggg 180gacatgagtc agtcta 196 <210> SEQ ID NO 209 <211> LENGTH: 345 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 53, 56 <223> OTHER INFORMATION: n = A,T,Cor G <400> SEQUENCE: 209 gacgcttggc cacttgacac cttttatttt ttaaggattcttaagtcatt tangtnactt 60 tgtaagtttt tcctgtgccc ccataagaat gatagctttaaaaattatgc tggggtagca 120 aagaagatac ttctagcttt agaatgtgta ggtatagccaggattcttgt gaggaggggt 180 gatttagagc aaatttctta ttctccttgc ctcatctgtaacatggggat aataatagaa 240 ctggcttgac aaggttggaa ttagtattac atggtaaatacatgtaaaat gtttagaatg 300 gtgccaagta tctaggaagt acttgggcat gggtggtaaatggct 345 <210> SEQ ID NO 210 <211> LENGTH: 178 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 210 gacgcttggc cacttgacactagagtaggg tttggccaac tttttctata aaggaccaga 60 gagtaaatat ttcaggctttgtgggttgtg cagtctctct tgcaactact cagctctgcc 120 attgtagcat agaaatcagccatagacagg acagaaatga atgggtggta aatggcta 178 <210> SEQ ID NO 211 <211>LENGTH: 454 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:211 tgggcacctt caatatctat ccagcgcatc taaattcgct tttttcttga ttaaaaattt 60caccacttgc tgtttttgct catgtatacc aagtagcagt ggtgtgaggc catgcttgtt 120ttttgattcg atatcagcac cgtataagag cagtgctttg gccattaatt tatcttcatt 180gtagacagca tagtgtagag tggtatctcc atactcatct ggaatatttg gatcagtgcc 240atgttccagc aacattaacg cacattcatc ttcctggcat tgtacggcct ttgtcagagc 300tgtcctcttt ttgttgtcaa ggacattaag ttgacatcgt ctgtccagca cgagttttac 360tacttctgaa ttcccattgg cagaggccag atgtagagca gtcctctttt gcttgtccct 420cttgttcaca tcagtgtccc tgagcataac ggaa 454 <210> SEQ ID NO 212 <211>LENGTH: 337 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:212 tccgttatgc cacccagaaa acctactgga gttacttatt aacatcaagg ctggaaccta 60tttgcctcag tcctatctga ttcatgagca catggttatt actgatcgca ttgaaaacat 120tgatcacctg ggtttcttta tttatcgact gtgtcatgac aaggaaactt acaaactgca 180acgcagagaa actattaaag gtattcagaa acgtgaagcc agcaattgtt tcgcaattcg 240gcattttgaa aacaaatttg ccgtggaaac tttaatttgt tcttgaacag tcaagaaaaa 300cattattgag gaaaattaat atcacagcat aacggaa 337 <210> SEQ ID NO 213 <211>LENGTH: 715 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 552, 630, 649, 657, 691,693, 697 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 213tcgggtgatg cctcctcagg catcttccat ccatctcttc aagattagct gtcccaaatg 60tttttccttc tcttctttac tgataaattt ggactccttc ttgacactga tgacagcttt 120agtatccttc ttgtcacctt gcagacttta aacataaaaa tactcattgg ttttaaaagg 180aaaaaagtat acattagcac tattaagctt ggccttgaaa cattttctat cttttattaa 240atgtcggtta gctgaacaga attcatttta caatgcagag tgagaaaaga agggagctat 300atgcatttga gaatgcaagc attgtcaaat aaacatttta aatgctttct taaagtgagc 360acatacagaa atacattaag atattagaaa gtgtttttgc ttgtgtacta ctaattaggg 420aagcaccttg tatagttcct cttctaaaat tgaagtagat tttaaaaacc catgtaattt 480aattgagctc tcagttcaga ttttaggaga attttaacag ggatttggtt ttgtctaaat 540tttgtcaatt tntttagtta atctgtataa ttttataaat gtcaaactgt atttagtccg 600ttttcatgct gctatgaaag aaatacccan gacagggtta tttataaang gaaagangtt 660aatttgactc ccagttcaca ggcctgagga ngnatcnccc gaaatcctta ttgcg 715 <210>SEQ ID NO 214 <211> LENGTH: 345 <212> TYPE: DNA <213> ORGANISM: Homosapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 6,8, 15 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 214ggtaangngc atacntcggt gctccggccg ccggagtcgg gggattcggg tgatgcctcc 60tcaggcccac ttgggcctgc ttttcccaaa tggcagctcc tctggacatg ccattccttc 120tcccacctgc ctgattcttc atatgttggg tgtccctgtt tttctggtgc tatttcctga 180ctgctgttca gctgccactg tcctgcaaag cctgcctttt taaatgcctc accattcctt 240catttgtttc ttaaatatgg gaagtgaaag tgccacctga ggccgggcac agtggctcac 300gcctgtaatc ccagcacttt gggagcctga ggaggcatca cccga 345 <210> SEQ ID NO215 <211> LENGTH: 429 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 215 ggtgatgcct cctcaggcga agctcaggga ggacagaaac ctcccgtggagcagaagggc 60 aaaagctcgc ttgatcttga ttttcagtac gaatacagac cgtgaaagcggggcctcacg 120 atccttctga ccttttgggt tttaagcagg aggtgtcaga aaagttaccacagggataac 180 tggcttgtgg cggccaagcg ttcatagcga cgtcgctttt tgatccttcgatgtcggctc 240 ttcctatcat tgtgaagcag aattcaccaa gcgttggatt gttcacccactaatagggaa 300 cgtgagctgg gtttagaccg tcgtgagaca ggttagtttt accctactgatgatgtgtkg 360 ttgccatggt aatcctgctc agtacgagag gaaccgcagg ttcasacatttggtgtatgt 420 gcttgcctt 429 <210> SEQ ID NO 216 <211> LENGTH: 593 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 15, 429, 446, 498, 512, 538, 543, 557 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 216 tgacacctatgtccngcatc tgttcacagt ttccacaaat agccagcctt tggccacctc 60 tctgtcctgaggtatacaag tatatcagga ggtgtatacc ttctcttctc ttccccacca 120 aagagaacatgcaggctctg gaagctgtct taggagcctt tgggctcaga atttcagagt 180 cttgggtaccttggatgtgg tctggaagga gaaacattgg ctctggataa ggagtacagc 240 cggaggagggtcacagagcc ctcagctcaa gcccctgtgc cttagtctaa aagcagcttt 300 ggatgaggaagcaggttaag taacatacgt aagcgtacac aggtagaaag tgctgggagt 360 cagaattgcacagtgtgtag gagtagtacc tcaatcaatg agggcaaatc aactgaaaga 420 agaagaccnattaatgaatt gcttangggg aaggatcaag gctatcatgg agatctttct 480 aggaagattattgtttanaa ttatgaaagg antagggcag ggacagggcc agaagtanaa 540 ganaacattgcctatanccc ttgtcttgca cccagatgct ggacaaggtg tca 593 <210> SEQ ID NO 217<211> LENGTH: 335 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 217 tgacaccttg tccagcatct gacgtgaaga tgagcagctc agaggaggtgtcctggattt 60 cctggttctg tgggctccgt ggcaatgaat tcttctgtga agtggatgaagactacatcc 120 aggacaaatt taatcttact ggactcaatg agcaggtccc tcactatcgacaagctctag 180 acatgatctt ggacctggag cctgatgaag aactggaaga caaccccaaccagagtgacc 240 tgattgagca ggcagccgag atgctttatg gattgatcca cgcccgctacatccttacca 300 accgtggcat cgcccagatg ctggacaagg tgtca 335 <210> SEQ IDNO 218 <211> LENGTH: 248 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 218 tacgtactgg tcttgaaggt cttaggtaga gaaaaaatgtgaatatttaa tcaaagacta 60 tgtatgaaat gggactgtaa gtacagaggg aagggtggcccttatcgcca gaagttggta 120 gatgcgtccc cgtcatgaaa tgttgtgtca ctgcccgacatttgccgaat tactgaaatt 180 ccgtagaatt agtgcaaatt ctaacgttgt tcatctaagattatggttcc atgtttctag 240 tactttta 248 <210> SEQ ID NO 219 <211> LENGTH:530 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 49, 216, 265, 275, 281, 296, 371,407, 424, 429, 454, 456, 458, 464, 474, 476, 506, 509, 527, 530 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 219 tgacgcttggccacttgaca caagtagggg ataaggacaa agacccatna ggtggcctgt 60 cagccttttgttactgttgc ttccctgtca ccacggcccc ctctgtaggg gtgtgctgtg 120 ctctgtggacattggtgcat tttcacacat accattctct ttctgcttca cagcagtcct 180 gaggcgggagcacacaggac taccttgtca gatgangata atgatgtctg gccaactcac 240 cccccaaccttctcactagt tatangaaga gccangccta naaccttcta tcctgncccc 300 ttgccctatgacctcatccc tgttccatgc cctattctga tttctggtga actttggagc 360 agcctggtttntcctcctca ctccagcctc tctccatacc atggtanggg ggtgctgttc 420 cacncaaanggtcaggtgtg tctggggaat cctnananct gccnggagtt tccnangcat 480 tcttaaaaaccttcttgcct aatcanatng tgtccagtgg ccaaccntcn 530 <210> SEQ ID NO 220<211> LENGTH: 531 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 220 tgacgcttgg ccacttgaca ctaaatagca tcttctaaag gcctgattcagagttgtgga 60 aaattctccc agtgtcaggg attgtcagga acagggctgc tcctgtgctcactttacctg 120 ctgtgtttct gctggaaaag gagggaagag gaatggctga tttttacctaatgtctccca 180 gtttttcata ttcttcttgg atcctcttct ctgacaactg ttcccttttggtcttcttct 240 tcttgctcag agagcaggtc tctttaaaac tgagaaggga gaatgagcaaatgattaaag 300 aaaacacact tctgaggccc agagatcaaa tattaggtaa atactaaaccgcttgcctgc 360 tgtggtcact tttctcctct ttcacatgct ctatccctct atcccccacctattcatatg 420 gcttttatct gccaagttat ccggcctctc atcaaccttc tcccctagcctactggggga 480 tatccatctg ggtctgtctc tggtgtattg gtgtcaagtg gccaagcgtc a531 <210> SEQ ID NO 221 <211> LENGTH: 530 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 221 attgacgctt ggccacttgacacccgcctg cctgcaatac tggggcaagg gccttcactg 60 ctttcctgcc accagctgccactgcacaca gagatcagaa atgctaccaa ccaagactgt 120 tggtcctcag cctctctgaggagaaagagc agaagcctgg aagtcagaag agaagctaga 180 tcggctacgg ccttggcagccagcttcccc acctgtggca ataaagtcgt gcatggctta 240 acaatggggg cacctcctgagaaacacatt gttaggcaat tcggcgtgtg ttcatcagag 300 catatttaca caaacctcgatagtgcagcc tactatccac tattgctcct acgctgcaaa 360 cctgaacagc atgggactgtactgaatact ggaagcagct ggtgatggta cttatttgtg 420 tatctaaaca cagagaaggtacagtaagaa tatggtatca taaacttaca gggaccgcca 480 tcctatatgc agtctgttgtgaccaaaatg tgtcaagtgg ccaagcgtca 530 <210> SEQ ID NO 222 <211> LENGTH:578 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 308, 381, 561, 570, 573 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 222 tgtatcgacgtagtggtctc cgggctacta ggccgttgtg tgctggtagt acctggttca 60 ctgaaaggcgcatctccctc cccgcgtcgc cctgaagcag ggggaggact tcgcccagcc 120 aaggcagttgtatgagtttt agctgcggca cttcgagacc tctgagccca cctccttcag 180 gagccttccccgattaagga agccagggta aggattcctt cctcccccag acaccacgaa 240 caaaccaccaccccccctat tctggcagcc catatacatc agaacgaaac aaaaataaca 300 aataaacnaaaaccaaaaaa aaaagagaag gggaaatgta tatgtctgtc catcctgttg 360 ctttagcctgtcagctccta nagggcaggg accgtgtctt ccgaatggtc tgtgcagcgc 420 cgactgcgggaagtatcgga ggaggaagca gagtcagcag aagttgaacg gtgggcccgg 480 cggctcttgggggctggtgt tgtacttcga gaccgctttc gctttttgtc ttagatttac 540 gtttgctctttggagtggga naccactacn tcnataca 578 <210> SEQ ID NO 223 <211> LENGTH: 578<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 223tgtatcgacg tagtggtctc ctcttgcaaa ggactggctg gtgaatggtt tccctgaatt 60atggacttac cctaaacata tcttatcatc attaccagtt gcaaaatatt agaatgtgtt 120gtcactgttt catttgattc ctagaaggtt agtcttagat atgttacttt aacctgtatg 180ctgtagtgct ttgaatgcat tttttgtttg catttttgtt tgcccaacct gtcaattata 240gctgcttagg tctggactgt cctggataaa gctgttaaaa tattcaccag tccagccatc 300ttacaagcta attaagtcaa ctaaatgctt ccttgttttg ccagacttgt tatgtcaatc 360ctcaatttct gggttcattt tgggtgccct aaatcttagg gtgtgacttt cttagcatcc 420tgtaacatcc attcccaagc aagcacaact tcacataata ctttccagaa gttcattgct 480gaagcctttc cttcacccag cggagcaact tgattttcta caacttccct catcagagcc 540acaagagtat gggatatgga gaccactacg tcgataca 578 <210> SEQ ID NO 224 <211>LENGTH: 345 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 13 <223> OTHER INFORMATION:n = A,T,C or G <400> SEQUENCE: 224 tgtatcgacg tantggtctc ccaaggtgctgggattgcag gcatgagcca ccactcccag 60 gtggatcttt ttctttatac ttacttcattaggtttctgt tattcaagaa gtgtagtggt 120 aaaagtcttt tcaatctaca tggttaaataatgatagcct gggaaataaa tagaaatttt 180 ttctttcatc tttaggttga ataaagaaacagaaaaaata gaacatactg aaaataatct 240 aagttccaac catagaagaa ctgcagaagaaatgaagaaa gtgatgatga tttagatttt 300 gatattgatt tagaagacac aggaggagaccactacgtcg ataca 345 <210> SEQ ID NO 225 <211> LENGTH: 347 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 225 tgtatcgacgtagtggtctc caaactgagg tatgtgtgcc actagcacac aaagccttcc 60 aacagggacgcaggcacagg cagtttaaag ggaatctgtt tctaaattaa tttccacctt 120 ctctaagtattctttcctaa aactgatcaa ggtgtgaagc ctgtgctctt tcccaactcc 180 cctttgacaacagccttcaa ctaacacaag aaaaggcatg tctgacactc ttcctgagtc 240 tgactctgatacgttgttct gatgtctaaa gagctccaga acaccaaagg gacaattcag 300 aatgctggtgtataacagac tccaatggag accactacgt cgataca 347 <210> SEQ ID NO 226 <211>LENGTH: 281 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 4, 6, 11 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 226 aggngnggga ntgtatcgacgtagtggtct cccaacagtc tgtcattcag tctgcaggtg 60 tcagtgtttt ggacaatgaggcaccattgt cacttattga ctcctcagct ctaaatgctg 120 aaattaaatc ttgtcatgacaagtctggaa ttcctgatga ggttttacaa agtattttgg 180 atcaatactc caacaaatcagaaagccaga aagaggatcc tttcaatatt gcagaaccac 240 gagtggattt acacacctcaggagaccact acgtcgatac a 281 <210> SEQ ID NO 227 <211> LENGTH: 3646 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 227 gggaaacacttcctcccagc cttgtaaggg ttggagccct ctccagtata tgctgcagaa 60 tttttctctcggtttctcag aggattatgg agtccgcctt aaaaaaggca agctctggac 120 actctgcaaagtagaatggc caaagtttgg agttgagtgg ccccttgaag ggtcactgaa 180 cctcacaattgttcaagctg tgtggcgggt tgttactgaa actcccggcc tccctgatca 240 gtttccctacattgatcaat ggctgagttt ggtcaggagc accccttccg tggctccact 300 catgcaccattcataatttt acctccaagg tcctcctgag ccagaccgtg ttttcgcctc 360 gaccctcagccggttcggct cgccctgtac tgcctctctc tgaagaagag gagagtctcc 420 ctcacccagtcccaccgcct taaaaccagc ctactccctt agggtcatcc catgtctcct 480 cggctatgtcccctgtaggc tcatcaccca ttgcctcttg gttgcaaccg tggtgggagg 540 aagtagcccctctactacca ctgagagagg cacaagtccc tctgggtgat gagtgctcca 600 cccccttcctggtttatgtc ccttctttct acttctgact tgtataattg gaaaacccat 660 aatcctcccttctctgaaaa gccccaggct ttgacctcac tgatggagtc tgtactctgg 720 acacattggcccacctggga tgactgtcaa cagctccttt tgaccctttt cacctctgaa 780 gagagggaaagtatccaaag agaggccaaa aagtacaacc tcacatcaac caataggccg 840 gaggaggaagctagaggaat agtgattaga gacccaattg ggacctaatt gggacccaaa 900 tttctcaagtggagggagaa cttttgacga tttccaccgg tatctcctcg tgggtattca 960 gggagctgctcagaaaccta taaacttgtc taaggcgact gaagtcgtcc aggggcatga 1020 tgagtcaccaggagtgtttt tagagcacct ccaggaggct tatcagattt acaccccttt 1080 tgacctggcagcccccgaaa atagccatgc tcttaatttg gcatttgtgg ctcaggcagc 1140 cccagatagtaaaaggaaac tccaaaaact agagggattt tgctggaatg aataccagtc 1200 agcttttagagatagcctaa aaggtttttg acagtcaaga ggttgaaaaa caaaaacaag 1260 cagctcaggcagctgaaaaa agccactgat aaagcatcct ggagtatcag agtttactgt 1320 tagatcagcctcatttgact tcccctccca catggtgttt aaatccagct acactacttc 1380 ctgactcaaactccactatt cctgttcatg actgtcagga actgttggaa actactgaaa 1440 ctggccgacctgatcttcaa aatgtgcccc taggaaaggt ggatgccacc atgttcacag 1500 acagtagcagcttcctcgag aagggactac gaaaggccgg tgcagctgtt accatggaga 1560 cagatgtgttgtgggctcag gctttaccag caaacacctc agcacaaaag gctgaattga 1620 tcgccctcactcaggctctc cgatggggta aggatattaa cgttaacact gacagcaggt 1680 acgcctttgctactgtgcat gtacgtggag ccatctacca ggagcgtggg ctactcacct 1740 cagcaggtggctgtaatcca ctgtaaagga catcaaaagg aaaacacggc tgttgcccgt 1800 ggtaaccagaaagctgattc agcagctcaa gatgcagtgt gactttcagt cacgcctcta 1860 aacttgctgcccacagtctc ctttccacag ccagatctgc ctgacaatcc cgcatactca 1920 acagaagaagaaaactggcc tcagaactca gagccaataa aaatcaggaa ggttggtgga 1980 ttcttcctgactctagaatc ttcatacccc gaactcttgg gaaaacttta atcagtcacc 2040 tacagtctaccacccattta ggaggagcaa agctacctca gctcctccgg agccgtttta 2100 agatcccccatcttcaaagc ctaacagatc aagcagctct ccggtgcaca acctgcgccc 2160 aggtaaatgccaaaaaaggt cctaaaccca gcccaggcca ccgtctccaa gaaaactcac 2220 caggagaaaagtgggaaatt gactttacag aagtaaaacc acaccgggct gggtacaaat 2280 accttctagtactggtagac accttctctg gatggactga agcatttgct accaaaaacg 2340 aaactgtcaatatggtagtt aagtttttac tcaatgaaat catccctcga catgggctgc 2400 ctgtttgccatagggtctga taatggaccg gccttcgcct tgtctatagt ttagtcagtc 2460 agtaaggcgttaaacattca atggaagctc cattgtgcct atcgacccca gagctctggg 2520 caagtagaacgcatgaactg caccctaaaa aacactctta caaaattaat cttagaaacc 2580 ggtgtaaattgtgtaagtct ccttccttta gccctactta gagtaaggtg caccccttac 2640 tgggctgggttcttaccttt tgaaatcatg tatgggaggg tgctgcctat cttgcctaag 2700 ctaagagatgcccaattggc aaaaatatca caaactaatt tattacagta cctacagtct 2760 ccccaacaggtacaagatat catcctgcca cttgttcgag gaacccatcc caatccaatt 2820 cctgaacagacagggccctg ccattcattc ccgccaggtg acctgttgtt tgttaaaaag 2880 ttccagagagaaggactccc tcctgcttgg aagagacctc acaccgtcat cacgatgcca 2940 acggctctgaaggtggatgg cattcctgcg tggattcatc actcccgcat caaaaaggcc 3000 aacagagcccaactagaaac atgggtcccc agggctgggt caggcccctt aaaactgcac 3060 ctaagttgggtgaagccatt agattaattc tttttcttaa ttttgtaaaa caatgcatag 3120 cttctgtcaaacttatgtat cttaagactc aatataaccc ccttgttata actgaggaat 3180 caatgatttgattcccccaa aaacacaagt ggggaatgta gtgtccaacc tggtttttac 3240 taaccctgtttttagactct ccctttcctt taatcactca gcttgtttcc acctgaattg 3300 actctcccttagctaagagc gccagatgga ctccatcttg gctctttcac tggcagccgc 3360 ttcctcaaggacttaacttg tgcaagctga ctcccagcac atccaagaat gcaattaact 3420 gataagatactgtggcaagc tatatccgca gttcccagga attcgtccaa ttgatcacag 3480 cccctctacccttcagcaac caccaccctg atcagtcagc agccatcagc accgaggcaa 3540 ggccctccaccagcaaaaag attctgactc actgaagact tggatgatca ttagtatttt 3600 tagcagtaaagttttttttt ctttttcttt ctttttttct cgtgcc 3646 <210> SEQ ID NO 228 <211>LENGTH: 419 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 402 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 228 taagagggta caagatctaagcacagccgt caatgcagaa cacagaacgt agcctggtaa 60 gtgtgttaag agtgggaatttttggagtac agagtaaggc acctaaccct agctggggtt 120 tggtgacggt cccagatggcttacagaaga aagtgtcctg agatgagttt ttaagaatga 180 ataaggatag acacaagtgaggactgactt ggcagtggtg aatggtgggt ggcaaaaaac 240 ttcgcatgta tggaaactgcacgtacagga atgaagaatg agactgtgtg gtgtttaatg 300 agctgcaaat actaattttatcctgaaagt tttgaagagt taactaaaaa gtatttttta 360 gtaaggaaat aaccctacatttcagggtta ttgtttgttt anatattgaa ggtgcccaa 419 <210> SEQ ID NO 229 <211>LENGTH: 148 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:229 aagagggtac ctgtatgtag ccatggtggc aatgagagac tgattactac ctgctggaga 60ttgtttaagt gagttaatat attaaggata aagggagcca ggttttttga ctgttggaga 120aggaaattac agatattgaa ggtcccaa 148 <210> SEQ ID NO 230 <211> LENGTH: 257<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 230taagagggta cmaaaaaaaa aaaatagaac gaatgagtaa gacctactat ttgatagtac 60aacagggtga ctatagtcaa tgataactta attatacatt taacatagag tgtaattgga 120ttgtttgtaa ctcgaaggat aaatgcttga gaggatggat accccattct ccatgatgta 180cttatttcac attacatgcc tgtatcaaag catctcatat accctataaa tatgtacacc 240tactatgtac cctctta 257 <210> SEQ ID NO 231 <211> LENGTH: 260 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 231 taagagggtacgggtatttg ctgatgggat ttttttttct ttctttttct ttggaaaaca 60 aaatgaaagccagaacaaaa ttattgaaca aaagacaggg actaaatctg gagaaatgaa 120 gtcccctcacctgactgcca tttcattcta tctgaccttc cagtctaggt taggagaata 180 gggggtggaggggattaatc tgatacaggt atatttaaag caactctgca tgtgtgccag 240 aagtccatggtaccctctta 260 <210> SEQ ID NO 232 <211> LENGTH: 596 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature<222> LOCATION: 437, 440, 461, 536, 541, 565, 580, 587, 590, 595 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 232 tgctcctcttgccttaccaa ccacaaatta gaaccataat gagatgtcac ctcatacctg 60 gtgggattaacattatttaa aaaatcagaa gtattgacaa ggatgtgaag aaattagaac 120 atctgtgcactgttggtggg aatgtaaaaa aggtgtggcc actatgggta acagcatgaa 180 ggttcctcaaaaaaaatttt ttttaatcta ctctatgatc gatcttgagg ttgtttatgc 240 aaaagaactgaaatcaggat tttgaggaaa tattcacatt cccacatcca tttctgcttt 300 attcataatactcaagagat ggaaacaacc taaatgtcca tcccgggatg aatggataaa 360 cacagtgtggtatatgcata caatggaata ttatttagtc tttaaaaaga aaaattctat 420 catatactacaacttanatn aaccttgagg acacaatgct nagtgaaata agccacggaa 480 ggacgaatactgcattattc ccttatatga agtatctaaa gtggtcaaac tcttanagca 540 naaagtaaaaatgggtggtt gccanacagt tggttaggcn agaaganaan cctant 596 <210> SEQ ID NO233 <211> LENGTH: 96 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 233 tcttctgaag acctttcgcg actcttaagc tcgtggttgg taaggcaagaggagcgttgg 60 taaggcaaga ggagcgttgg taaggcaaga ggagca 96 <210> SEQ ID NO234 <211> LENGTH: 313 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 234 tgtaagtcga gcagtgtgat gataaaactt gaatggatca atagttgcttcttatggatg 60 agcaaagaaa gtagtttctt gtgatggaat ctgctcctgg caaaaatgctgtgaacgttg 120 ttgaaaagac aacaaagagt ttagagtagt acataaattt agaatagtacataaacttag 180 aatagtacat aaacttagta cataaataat gcacgaagca ggggcagggcttgagagaat 240 tgacttcaat ttggaaagag tatctactgt aggttagatg ctctcaaacagcatcacact 300 gctcgactta caa 313 <210> SEQ ID NO 235 <211> LENGTH: 550<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 235aacgaggaca gatccttaaa aagaatgttg agtgaaaaaa gtagaaaata agataatctc 60caaagtccag tagcattatt taaacatttt taaaaaatac actgataaaa attttgtaca 120tttcccaaaa atacatatgg aagcacagca gcatgaatgc ctatgggrtt gaggataggg 180gttgggagta gggatgggga taaaggggga aaataaaacc agagaggagt cttacacatt 240tcatgaacca aggagtataa ttatttcaac tatttgtacc wgaagtccag aaagagtgga 300ggcagaaggg ggagaagagg gcgaagaaac gtttttggga gaggggtccc asaagagaga 360ttttcgcgat gtggcgctac atacgttttt ccaggatgcc ttaagctctg caccctattt 420ttctcatcac taatattaga ttaaaccctt tgaagacagc gtctgtggtt tctctacttc 480agctttccct ccgtgtcttg cacacagtag ctgttttaca agggttgaac tgactgaagt 540gagattattc 550 <210> SEQ ID NO 236 <211> LENGTH: 325 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 236 tagactgact catgtcccctaccagagtag ctagaattaa tagcacaagc ctctacaccc 60 aggaactcac tattgaatacataaatggaa tttattcagc cttaaaaagt ttggaaggaa 120 attctgacat atgctaaaacatggatgaac cttgaagact ttatgataag taaaagaagc 180 cagtcataaa aggaaaaatattgcatgatt ccacttatat gaggtaccta gagtagtcaa 240 tttcatagaa acacaaaatagaatggtgtt tgccagggct tttgaggaaa agggaatgac 300 aagttagggg acatgagtcagtcta 325 <210> SEQ ID NO 237 <211> LENGTH: 373 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 355 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE:237 tagactgact catgtcccct atctactcaa catttccact tgaagtctga taggcatctc 60agacttatct tgtcccaaag caaactcttt atttcttttc atcctagtct ttatttcttg 120tgctgtctta cccatctcaa aagagtgcca aaatccacca agttgctgaa acagaaatct 180aagaaatatc cttgattctt ctttttccca tctacttcac ttctaattca ttagtaaata 240atctgtttca gaaaaccaaa cacctcatgt tctcactcat aagggggagt tgaacaatga 300gaacacacag acacagggag gggaacatca cacaccacgg cccgtcaggg agtangggac 360atgagtcagt cta 373 <210> SEQ ID NO 238 <211> LENGTH: 492 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature<222> LOCATION: 272, 310, 380, 435, 474, 484, 488 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 238 tagactgact catgtcccctataatgctcc caggcatcag aaagcatctc aaactggagc 60 tgacaccatg gcagaggtttcaggtaagtc acaaaagggg tcctaaagaa tttgccctca 120 atatcagagt gattagaagaagtggacaga gctacccaag ttaaacatat gcgagataaa 180 aaaaatatgg cacttgtgaacacacactac aggaggaaaa taaggaacat aatagcatat 240 tgtgctatta tgatgatgaagaacctctct anaagaaaac ataaccaaag aaacaaagaa 300 aattcctgcn aatgtttaatgctatagaag aaattaacaa aaacatatat tcaatgaatt 360 cagaaaagtt agcaggtcanaagaaaacaa atcaaagacc agaataatcc cattttagat 420 tgtcgagtaa actanaacagaaagaatacc actggaaatt gaattcctac gtangggaca 480 tgantcantc ta 492 <210>SEQ ID NO 239 <211> LENGTH: 482 <212> TYPE: DNA <213> ORGANISM: Homosapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 245<223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 239 tggaaagtatttaatgatgg gcaacttgct gtttacttcc tacatatccc atcatcttct 60 gtatttttttaaataacttt tttttggatt tttaaagtaa ccttattctg agaggtaaca 120 tggattacatacttctaagc cattaggaga ctctatgtta aaccaaaagg aaatgttact 180 agatcttcatttgatcaata ggatgtgata atcatcatct ttctgctcta atggaaaagt 240 actanaaacatggaaccata atcttagatg aacaacgtta gaatttgcac taattctacg 300 gaatttcagtaattcggcaa atgtcgggca gtgacacaac atttcatgac ggggacgcat 360 ctaccaacttctggcgataa gggccaccct tccctctgta cttacagtcc catttcatac 420 acagtctttgattaaatatt cacatttttt ctctacctaa agaccttcaa gaccagtacg 480 ta 482 <210>SEQ ID NO 240 <211> LENGTH: 519 <212> TYPE: DNA <213> ORGANISM: Homosapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 491<223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 240 tgtatcgacgtagtggtctc cccatgtgat agtctgaaat atagcctcat gggatgagag 60 gctgtgccccagcccgacac ccgtaaaggg tctgtgctga ggtggattag taaaagagga 120 aagccttgcagttgagatag aggaagggca ctgtctcctg cctgcccctg ggaactgaat 180 gtctcggtataaaacccgat tgtacatttg ttcaattctg agataggaga aaaaccaccc 240 tatggcgggaggcgagacat gttggcagca atgctgcctt gttatgcttt actccacaga 300 tgtttgggcggagggaaaca taaatctggc ctacgtgcac atccaggcat agtacctccc 360 tttgaacttaattatgacac agattccttt gctcacatgt ttttttgctg accttctcct 420 tattatcaccctgctctcct accgcattcc ttgtgctgag ataatgaaaa taatatcaat 480 aaaaacttganggaactcgg agaccactac gtcgataca 519 <210> SEQ ID NO 241 <211> LENGTH:771 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 304, 402, 442, 463, 510, 541,550, 567, 571, 596, 617, 624, 644, 648, 652, 667, 682, 686, 719, 722,729, 732, 751, 752, 757, 758, 760, 763, 766, 769 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 241 tgtatcgacg tagtggtctccactcccgcc ttgacggggc tgctatctgc cttccaggcc 60 actgtcacgg ctcccgggtagaagtcactt atgagacaca ccagtgtggc cttgttggct 120 tgaagctcct cagaggagggtgggaacaga gtgaccgagg gggcagcctt gggctgacct 180 aggacggtca gcttggtccctccgccaaac acgagagtgc tgctgcttgt atatgagctg 240 cagtaataat cagcctcgtcctcagcctgg agcccagaga tggtcaggga ggccgtgttg 300 ccanacttgg agccagagaagcgattagaa acccctgagg gccgattacc gacctcataa 360 atcatgaatt tgggggctttgcctgggtgc tgttggtacc angagacatt attataacca 420 ccaacgtcac tgctggttccantgcaggga aaatggttga tcnaactgtc caagaaaacc 480 actacgtcca taccaatccactaattgccn gccgcctgca ggttcaacca tattggggaa 540 naactccccn ccgccgtttgggattgncat naacctttga aattttttcc tattanttgt 600 ccccctaaaa taaaccnttgggcnttaatc cattgggtcc atancttntt tncccggttt 660 ttaaaanttg tttatcccgccncccnattt cccccccaac tttccaaaac ccgaaaccnt 720 tnaaatttnt tnaaaccctggggggttccc nnaattnnan ttnaanctnc c 771 <210> SEQ ID NO 242 <211> LENGTH:167 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 242tgggcacctt caatatcggg ctcatcgata acatcacgct gctgatgctg ctgttgctgg 60tcctctctag gaacctctgg attttcaaat tctttgagga attcatccaa attatctgcc 120tctcctcctt tcctcctttt tctaaggtct tctggtacaa gcggtca 167 <210> SEQ ID NO243 <211> LENGTH: 338 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 243 ttgggcacct tcaatatcta ctgatctaaa tagtgtggtt tgaggcctcttgttcctggc 60 taaaaatcct tggcaagagt caatctccac tttacaatag aggtaaaaatcttacaatgg 120 atattcttga caaagctagc atagagacag caattttaca caaggtatttttcacctgtt 180 taataacagt ggttttccta cacccatagg gtgccaccaa gggaggagtgcacagttgca 240 gaaacaaatt aagatactga agacaacact acttaccatt tcccgtatagctaaccacca 300 gttcaactgt acatgtatgt tcttatgggc aatcaaga 338 <210> SEQID NO 244 <211> LENGTH: 346 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 244 tttttggctc ccatacagca cactctcatg ggaaatgtctgttctaaggt caacccataa 60 tgcaaaaatc atcaatatac ttgaagatcc ccgtgtaaggtacaatgtat ttaatattat 120 cactgataca attgatccaa taccagtttt agtctggcattgaatcaaat cactgttttt 180 gttgtataaa aagagaaata tttagcttat atttaagtaccatattgtaa gaaaaaagat 240 gcttatcttt acatgctaaa atcatgatct gtacattggtgcagtgaata ttactgtaaa 300 agggaagaag gaatgaagac gagctaagga tattgaaggtgcccaa 346 <210> SEQ ID NO 245 <211> LENGTH: 521 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 252, 337, 434, 455, 466, 478, 494, 510, 516 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 245 accaatccca cacggatactgagggacaag tatatcatcc catttcatcc ctacagcagc 60 aacttcatga ggcaggagttattagtccca ttttacagaa gaggaaactg agacttaggg 120 agatcaagta atttgcccaggtcgcacaat tagtgataga gccagggctt gaagcgacgt 180 ctgtcttaag ccaatgacccctgcagatta ttagagcaac tgttctccac aacagtgtaa 240 gcctcttgct anaagctcaggtccacaagg gcagagattt ttgtctgttt tgctcattgc 300 tccttcccca ttgcttagagcagggtctgc cacgaancag gttctcaatg catagttatt 360 aaatgtatat aagagcaaacatatgttaca gagaactttc tgtatgcttg tcacttacat 420 gaatcacctg tganatgggtatgcttgttc cccantgttg cagatnaaga tattgaangt 480 gcccaaatca ctanttgcgggcgcctgcan gtccancata t 521 <210> SEQ ID NO 246 <211> LENGTH: 482 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 464 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 246 tggaaccaat ccaaataccc atcaatgata gactggataaagaaaatttg gcacatgttc 60 accatgaaat actatgcagc cataaaaaag gatgagttcatatcctttgc agggacatgg 120 atgaagctgg agaccatcat tctcagcaaa ctaacaagggaacagaaaac caaacactgc 180 atgttctcac tcttaagtgg gagctgaaca atgagaacacatggacacag ggaggggaac 240 atcacacagt ggggcctgct ggtgggtagg ggtctaggggagggatagca ttaggagaaa 300 tacctaatgt agatgacggg ttgatgggtg cagcaaaccaccatgacacg tgtataccta 360 tgtaacaaac ctgcatgttc tgcacatgta ccccagaacttaaagtgtta ataaaaaaat 420 taagaaaaaa gttaagtatg tcatagatac ataaaatattgtanatattg aaggtgccca 480 aa 482 <210> SEQ ID NO 247 <211> LENGTH: 474<212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 220, 255, 287, 312, 339, 374,382, 403, 414, 426, 427, 428, 432, 433, 434, 435, 436, 465 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 247 ttcgatacag gcacagagtaagcagaaaaa tggctgtggt ttaaccaagt gagtacagtt 60 aagtgagaga ggggcagagaagacaagggc atatgcaggg ggtgattata acaggtggtt 120 gtgctgggaa gtgagggtactcggggatga ggaacagtga aaaagtggca aaaagtggta 180 agatcagtga attgtacttctccagaattt gatttctggn ggagtcaaat aactatccag 240 tttggggtat catanggcaacagttgaggt ataggaggta gaagtcncag tgggataatt 300 gaggttatga anggtttggtactgactggt actgacaang tctgggttat gaccatggga 360 atgaatgact gtanaagcgtanaggatgaa actattccac ganaaagggg tccnaaaact 420 aaaaannnaa gnnnnnggggaatattattt atgtggatat tgaangtgcc caaa 474 <210> SEQ ID NO 248 <211>LENGTH: 355 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 69, 87, 186, 192, 220, 227,251, 278, 339, 346, 350 <223> OTHER INFORMATION: n = A,T,C or G <400>SEQUENCE: 248 ttcgatacag gcaaacatga actgcaggag ggtggtgacg atcatgatgttgccgatggt 60 ccggatggnc acgaagacgc actggancac gtgcttacgt ccttttgctctgttgatggc 120 cctgagggga cgcaggaccc ttatgaccct cagaatcttc acaacgggagatggcactgg 180 attgantccc antgacacca gagacacccc aaccaccagn atatcantatattgatgtag 240 ttcctgtaga nggccccctt gtggaggaaa gctccatnag ttggtcatcttcaacaggat 300 ctcaacagtt tccgatggct gtgatgggca tagtcatant taaccntgtntcgaa 355 <210> SEQ ID NO 249 <211> LENGTH: 434 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 249 ttggattggt cctccaggagaacaagggga aaaaggtgac cgagggctcc ctggaactca 60 aggatctcca ggagcaaaaggggatggggg aattcctggt cctgctggtc ccttaggtcc 120 acctggtcct ccaggcttaccaggtcctca aggcccaaag ggtaacaaag gctctactgg 180 acccgctggc cagaaaggtgacagtggtct tccagggcct cctgggcctc caggtccacc 240 tggtgaagtc attcagcctttaccaatctt gtcctccaaa aaaacgagaa gacatactga 300 aggcatgcaa gcagatgcagatgataatat tcttgattac tcggatggaa tggaagaaat 360 atttggttcc ctcaattccctgaaacaaga catcgagcat atgaaatttc caatgggtac 420 tcagaccaat ccaa 434<210> SEQ ID NO 250 <211> LENGTH: 430 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:301, 430 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 250tggattggtc acatggcaga gacaggattc caaggcagtg agaggaggat acaatgcttc 60tcactagtta ttattattta ttttattttt gagatgaagt ctcgctttgt ctcccaggct 120ggagagcggt ggtgcgatct tggctctctg caacccccgc ctcaagcaat tctcctgtct 180tagcctcgcg ggtagatgga attacaggcg cccaccgcca tgcccaacta atttttttgt 240gtcttcagta gagacagggt ttcgccatgt tgggcaggct ggtcttgaac tcctgacctc 300nagtgatctg ccctcctcgg cctcacaaag tgctggaatt acaggcatgg gctgctgcac 360ccagtcaact tctcactagt tatggcctta tcattttcac cacattctat tggcccaaaa 420aaaaaaaaan 430 <210> SEQ ID NO 251 <211> LENGTH: 329 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 251 tggtactcca ccatyatggggtcaaccgcc atcctcgccc tcctcctggc tgttctccaa 60 ggagtctgtg ccgaggtgcagctgrtgcag tctggagcag aggtgaaaaa gtccggggag 120 tctctgaaga tctcctgtaagggttctgga tacaccttta agatctactg gatcgcctgg 180 gtgcgccagt tgcccgggaaaggcctggag tggatggggc tcatctttcc tgatgactct 240 gataccagat acagcccgtccttccaaggc caggtcacca tctcagtcga taagtccatc 300 agcaccgcct atctgcagtggagtaccaa 329 <210> SEQ ID NO 252 <211> LENGTH: 536 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 252 tggtactcca ctcagcccaaccttaattaa gaattaagag ggaacctatt actattctcc 60 caggctcctc tgctctaaccaggcttctgg gacagtatta gaaaaggatg tctcaacaag 120 tatgtagatc ctgtactggcctaagaagtt aaactgagaa tagcataaat cagaccaaac 180 ttaatggtcg ttgagacttgtgtcctggag cagctgggat aggaaaactt ttgggcagca 240 agaggaagaa ctgcctggaagggggcatca tgttaaaaat tacaagggga acccacacca 300 ggcccccttc ccagctctcagcctagagta ttagcatttc tcagctagag actcacaact 360 tccttgctta gaatgtgccaccggggggag tccctgtggg tgatgaggct ctcaagagtg 420 agagtggcat cctatcttctgtgtgcccac aggagcctgg cccgagactt agcaggtgaa 480 gtttctggtc caggctttgcccttgactca ctatgtgacc tctggtggag taccaa 536 <210> SEQ ID NO 253 <211>LENGTH: 507 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 1 <223> OTHER INFORMATION:n = A,T,C or G <400> SEQUENCE: 253 ntgttgcgat cccagtaact cgggaagctgaggcgggagg atcacctgag ctcaggaggt 60 tgaggccgca gtgagccggg accacgccactacactccag cctggggcat agagtgagac 120 cctccaagac agaaaagaaa agaaaggaagggaaagggaa agggaaaagg aaaaggaaaa 180 ggaaaaggaa aaggaaaaga caagacaaaacaagacttga atttggatct cctgacttca 240 attttatgtt ctttctacac cacaattcctctgcttacta agatgataat ttagaaaccc 300 ctcgttccat tctttacagc aagctggaagtttggtcaag taattacaat aatagtaaca 360 aatttgaata ttatatgcca ggtgtttttcattcctgctc tcacttaatt ctcaccactc 420 tgatataaat acaattgctg ccgggtgtggtggctcatgc ctgtaatccc ggcactttgg 480 gagaccgagg tgggcggats gcaacaa 507<210> SEQ ID NO 254 <211> LENGTH: 222 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:167 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 254ttggattggt cactgtgagg aagccaaatc ggatccgaga gtctttttct aaaggccagt 60actggccaca ctttctcctg ccgccttcct caaagctgaa gacacacaga gcaaggcgct 120tctgttttac tccccaatgg taactccaaa ccatagatgg ttagctnccc tgctcatctt 180tccacatccc tgctattcag tatagtccgt ggaccaatcc aa 222 <210> SEQ ID NO 255<211> LENGTH: 463 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 255 tgttgcgatc cataaatgct gaaatggaaa taaacaacat gatgagggaggattaagttg 60 gggagggagc acattaaggt ggccatgaag tttgttggaa gaagtgacttttgaacaagg 120 ccttggtgtt aagagctgat gagagtgtcc cagacagagg ggccactggtacaatagacg 180 agatgggaga gggcttggaa ggtgtgcgaa ataggaagga gtttgttctggtatgagtct 240 agtgaacaca gaggcgagag gccctggtgg gtgcagctgg agagttatgcagaataacat 300 taggccctgt gggggactgt agactgtcag caataatcca cagtttggattttattctaa 360 gagtgatggg aagccgtgga aagggggtta agcaaggagt gaaattatcagatttacagt 420 gataaaaata aattggtctg gctactgggg aaaaaaaaaa aaa 463 <210>SEQ ID NO 256 <211> LENGTH: 262 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 256 ttggattggt caacctgctc aactctacyt ttcctccttcttcctaaaaa attaatgaat 60 ccaatacatt aatgccaaaa cccttgggtt ttatcaatatttctgttaaa aagtattatc 120 cagaactgga cataatacta cataataata cataacaaccccttcatctg gatgcaaaca 180 tctattaata tagcttaaga tcactttcac tttacagaagcaacatcctg ttgatgttat 240 tttgatgttt ggaccaatcc aa 262 <210> SEQ ID NO257 <211> LENGTH: 461 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 2, 5, 6, 7, 8, 9,10, 11, 12, 13, 25, 32, 38, 71, 72 <223> OTHER INFORMATION: n = A,T,C orG <400> SEQUENCE: 257 gnggnnnnnn nnncaattcg actcngttcc cntggtanccggtcgacatg gccgcgggat 60 taccgcttgt nnctgggggt gtatggggga ctatgaccgcttgtagctgg gggtgtatgg 120 gggactatga ccgcttgtag mtggkggtgt atgggggactatgaccgctt gtcgggtggt 180 cggataaacc gacgcaaggg acgtgatcga agctgcgttcccgctctttc gcatcggtag 240 ggatcatgga cagcaatatc cgcattcgyc tgaaggcgttcgaccatcgc gtgctcgatc 300 aggcgaccgg cgacatcgcc gacaccgcac gccgtaccggcgcgctcatc cgcggtccga 360 tcccgcttcc cacgcgcatc gagaagttca cggtcaaccgtggcccgcac gtcgacaaga 420 agtcgcgcga gcagttcgag gtgcgtacct acaagcggtc a461 <210> SEQ ID NO 258 <211> LENGTH: 332 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 251 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE:258 tgaccgcttg tagctggggg tgtatggggg actacgaccg cttgtagctg ggggtgtatg 60ggggactatg accgcttgta gctgggggtg tatgggggac tatgaccgct tgtagctggg 120ggtgtatggg ggactaggac cgcttgtagc tgggggtgta tgggggacta tgaccgcttg 180tagctggggg tgtatggggg actacgaccg cttgtagctg ggggtgtatg ggggactatg 240accgcttgta nctgggggtg tatgggggac tatgaccgct tgtgctgcct gggggatggg 300aggagagttg tggttgggga aaaaaaaaaa aa 332 <210> SEQ ID NO 259 <211>LENGTH: 291 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 141, 144, 167, 168, 171,175, 194, 201, 202, 205, 209, 212, 235, 236, 245, 246, 258, 266, 268,270, 273, 277, 285, 290 <223> OTHER INFORMATION: n = A,T,C or G <400>SEQUENCE: 259 taccgcttgt gaccgcttgt gaccgcttgt gaccgcttgt gaccgcttgtgaccgcttgt 60 gaccgcttgt gaccgcttgt gaccgcttgt gaccgcttgt gaccgcttgtgaccgcttgt 120 gaccgcttgt gaccgcttgt nacngggggt gtctggggga ctatganngantgtnactgg 180 gggtgtctgg gggnctatga nngantgtna cngggggtgt ctgggggactatganngact 240 gtgcnncctg ggggatcnga ggagantngn ggntagngat ggttngggan a291 <210> SEQ ID NO 260 <211> LENGTH: 238 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 260 taagagggta ctggttaaaatacaggaaat ctggggtaat gaggcagaga accaggatac 60 tttgaggtca gggatgaaaactagaatttt tttctttttt tttgcctgag aaacttgctg 120 ctctgaagag gcccatgtattaattgcttt gatcttcctt ttcttacagc cctttcaagg 180 gcagagccct ccttatcctgaaggaatctt atccttagct atagtatgta ccctctta 238 <210> SEQ ID NO 261 <211>LENGTH: 746 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 662, 680, 685, 698, 707,709, 734, 740, 741 <223> OTHER INFORMATION: n = A,T,C or G <400>SEQUENCE: 261 ttgggcacct tcaatatcaa tagctaacat ttattgagtg tttatcgtatcataaaacac 60 tgttctaagc ctttaaacgt actaattcat ttaatgctca taatcactttagaaggtggg 120 tactagtatt agtctcattt acagatgcaa catgcaggca cagagaggttaattaacttg 180 cccaaggtaa cacagctaag aaatagaaaa aatattgaat ctggaaagttgggcttctgg 240 gtaacccaca gagtcttcaa tgagcctggg gcctcactca gtttgcttttacaaagcgaa 300 tgagtaacat cacttaattc agtgagtagg ccaaatggag gtcagctacgagtttctgct 360 gttcttgcag tggactgaca gatgtttaca acgtctggcc atcagtwaatggactgatta 420 tcattgggaw gtgggtgggc tgaatgttgg ccagtgaagt ttattcawgccatattttta 480 tgtttaggat gacttttggc tggtcctagg gcaagctctg tctgscacggaacacagaat 540 wacacaggga ccccctcaat ttctggtgtg gctagaacca tgaaccactggttgggggaa 600 caagcggtca aaacctaagt gcggccggct ggcagggtcc acccatatggggaaaactcc 660 cnacgcgttt ggaatgcctn agctngaatt attctaanag ttgtccncntaaaattagcc 720 tgggcgttaa tcangggtcn naagcc 746 <210> SEQ ID NO 262<211> LENGTH: 588 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 485, 488, 489,492, 493, 494, 496, 497, 498, 499, 502, 503, 504, 506, 521, 537, 550,564 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 262tgaccgcttg tcatctcaca tggggtcctg cacgcttttg cctttgtagg aaacctgaca 60tttgtctgtt tcttctttct cttttccttc ccatatcctc ctaatttacg tttgacttgt 120ttgctgagga ggcaggagct agagactgct gtgagctcat aggggtggga agtttatcct 180tcaagtcccg cccactcatc actgcttctc accttcccct gaccaggctt acaagtgggt 240tcttgcctgc tttccctttg gacccaacaa gcccctgtaa tgagtgtgca tgactctgac 300agctgtggac tcagggtcct tggctacagc tgccatgtaa aatatctcat ccagttctcg 360caaattgtta aaataaccac atttcttaga ttccagtacc caaatcatgt ctttacgaac 420tgctcctcac acccagaagt ggcacaataa ttcttgggga attattactt ttttttttct 480ctctnttnnc gnnngnnnng gnnngnccag gaattaccac nttggaagac ctggccngaa 540tttattatan aggggagccg attntttttc ctaacacaaa gcgggtca 588 <210> SEQ ID NO263 <211> LENGTH: 730 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 124, 510, 534,559, 604, 605, 635, 711, 729 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 263 tttttttttt tttggcctga gcaactgaaa ttatgaaatttccatatact caaaagagta 60 agactgcaaa aagattaaat gtaaaagttg tcttgtatacagtaatgttt aagataccta 120 ttanatttat aaatggaaaa ttagggcatt tggatatacaagttgaaaat tcaggagtga 180 ggttgggctg gctgggtata tactgaaaac tgtcagtacacagatgacat ctaaaaccac 240 aaatctggtt ttattttagc agtgatatgt gtcactcccacaaaagcctt cccaattggc 300 ctcagcatac acaacaagtc acctccccac agccctctacacataaacaa attccttagt 360 ttagttcagg aggaaatgcg cccttttcct tccgctctaggtgaccgcaa ggcccagttc 420 tcgtcaccaa gatgttaagg gaagtctgcc aaagaggcatctgaaaggaa ataaggggaa 480 tgggagtgac cacaaaggaa agccaaggan aaactttggagaccgtttct aganccctgg 540 catttcacaa caaaactcng gaacaaacct tgtctcatcaatcatttaag cccttcgttt 600 ggannagact ttctgaactg ggcgctgaac ataancctcattgaatgtct tcacagtctc 660 ccagctgaag gcacaccttg ggccagaagg ggaatcttccaggtcctcaa nacagggctc 720 gccctttgnc 730 <210> SEQ ID NO 264 <211>LENGTH: 715 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 364, 451, 476, 494, 495,515, 519, 524, 633, 635, 636, 645, 647, 649, 657, 692, 695, 701, 707,710, 713 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 264tttttttttt tttggccagt atgatagtct ctaccactat attgaagctc ttaggtcatt 60tacacttaat gtggttatag atgctgttga gcttacttct accaccttgc tatttctccc 120gtctcttttt tgttcctttt ctcttctttt cctcccttat tttataattg aattttttag 180gattctattt tatatagatt tatcagctat aacactttgt attcttttgt tttgtggttc 240ttctgtcatt tcaatgtgca tcttaaactc atcacaatct attttcaaat aatatcatat 300aaccttacat ataatgtaag aatctaccac catatatttc catttctccc ttccatccta 360tgtntgtcat attttttcct ttatatatgt tttaaagaca taatagtata tgggaggttt 420ttgcttaaaa tgtgatcaat attccttcaa ngaaacgtaa aaattcaaaa taaatntctg 480tttattctca aatnnaccta atatttccta ccatntctna tacntttcaa gaatctgaag 540gcattggttt tttccggctt aagaacctcc tctaaagcac tctaagcaga attaagtctt 600ctgggagagg aattctccca agcttgggcc ttnanntgta ctccntnang gttaaanttt 660ggccgggaaa tagaaattcc aagttaacag gntanttttt ntttttnttn tcncc 715 <210>SEQ ID NO 265 <211> LENGTH: 152 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 265 tttttttttt tttcccaaca caaagcacca ttatctttcctcacaatttt caacatagtt 60 tgattcccat gaagaggtta tgatttctaa agaaaacatggctactatac tatcaatcag 120 ggttaaatct tttttttttg agacggagtt ta 152 <210>SEQ ID NO 266 <211> LENGTH: 193 <212> TYPE: DNA <213> ORGANISM: Homosapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 180<223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 266 taaactccgtccccttctta atcaatatgg aggctaccca ctccacatta ccttcttttc 60 aagggactgtttccgtaact gttgtgggta ttcacgacca ggcttctaaa cctcttaaaa 120 ctccccaattctggtgccaa cttggacaac atgctttttt tttttttttt tttttttttn 180 gagacggagttta 193 <210> SEQ ID NO 267 <211> LENGTH: 460 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 267 tgttgcgatc ccttaagcatgggtgctatt aaaaaaatgg tggagaagaa aatacctgga 60 atttacgtct tatctttagagattgggaag accctgatgg aggacgtgga gaacagcttc 120 ttcttgaatg tcaattcccaagtaacaaca gtgtgtcagg cacttgctaa ggatcctaaa 180 ttgcagcaag gctacaatgctatgggattc tcccagggag gccaatttct gagggcagtg 240 gctcagagat gcccttcacctcccatgatc aatctgatct cggttggggg acaacatcaa 300 ggtgtttttg gactccctcgatgcccagga gagagctctc acatctgtga cttcatccga 360 aaaacactga atgctggggcgtactccaaa gttgttcagg aacgcctcgt gcaagccgaa 420 tactggcatg acccataaaaggaggatgtg gatcgcaaca 460 <210> SEQ ID NO 268 <211> LENGTH: 533 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 450, 470 <223> OTHER INFORMATION: n = A,T,Cor G <400> SEQUENCE: 268 tgttgcgatc cgttgataga atagcgacgt ggtaatgagtgcatggcacg cctccgactt 60 accttcgccc gtggggaccc cgagtacgtc tacggcgtcgtcacttagag taccctctgg 120 acgcccgggc gcgttcgatt taccggaagc gcgagctgcagtgggcttgc gcccccggcc 180 aaattctttg gggggtttaa ggccgcgggg aatttgaggtatctctatca gtatgtagcc 240 aagttggaac agtcgccatt cccgaaatcg ctttctttgaatccgcaccg cctccagcat 300 tgcctcattc atcaacctga aggcacgcat aagtgacggttgtgtcttca gcagctccac 360 tccataacta gcgcgctcga cctcgtcttc gtacgcgccaggtccgtgcg tgcgaattcc 420 caactccggt gagttgcgca tttcaagttn cgaaactgttcgcctccacn atttggcatg 480 ttcacgcatg acacggaata aactcgtcca gtaccgggaatgggatcgca aca 533 <210> SEQ ID NO 269 <211> LENGTH: 50 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 269 tttttttttt ttcgcctgaattagctacag atcctcctca caagcggtca 50 <210> SEQ ID NO 270 <211> LENGTH:519 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 270tgttgcgatc caaataaccc accagcttct tgcacacttc gcagaagcca ccgtcctttg 60gctgagtcac gtgaacggtc agtgcaagca gccgcgtgcc agagcagagg tgcagcatgc 120tgcacaccag ctcagggctg acctcctcca gcaggatgga caggatggag ctgccgtacg 180tgtccaccac ctcctggcac tcttccgaca gggacttcgg cagcttcgag cacattttgt 240caaaagcgtc gagtatttct ttctcagtct tgttgttgtc aatcagcttg gtcacctcct 300tcaccaggaa ttcacacacc tcacagtaaa catcagactt tgctgggacc tcgtgcttct 360taatgggctc caccagttcc agggcaggga tgacattctt ggaggccact ttggcgggga 420ccagagtctg catgggcatc tctttcacct catcacagaa cccaaccagc gcacagatct 480ccttgggttg catgtgcatc atcatctggg atcgcaaca 519 <210> SEQ ID NO 271 <211>LENGTH: 457 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:271 tttttttttt ttcgggcggc gaccggacgt gcactcctcc agtagcggct gcacgtcgtg 60ccaatggccc gctatgagga ggtgagcgtg tccggcttcg aggagttcca ccgggccgtg 120gaacagcaca atggcaagac cattttcgcc tactttacgg gttctaagga cgccgggggg 180aaaagctggt gccccgactg cgtgcaggct gaaccagtcg tacgagaggg gctgaagcac 240attagtgaag gatgtgtgtt catctactgc caagtaggag aagagcctta ttggaaagat 300ccaaataatg acttcagaaa aaacttgaaa gtaacagcag tgcctacact acttaagtat 360ggaacacctc aaaaactggt agaatctgag tgtcttcagg ccaacctggt ggaaatgttg 420ttctctgaag attaagattt taggatggca atcaaga 457 <210> SEQ ID NO 272 <211>LENGTH: 102 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:272 tttttttttt ttgggcaaca acctgaatac cttttcaagg ctctggcttg ggctcaagcc 60cgcaggggaa atgcaactgg ccaggtcaca gggcaatcaa ga 102 <210> SEQ ID NO 273<211> LENGTH: 455 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 380, 415, 454<223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 273 ttttttttttttggcaatca acaggtttaa gtcttcggcc gaagttaatc tcgtgttttt 60 ggcaatcaacaggtttaagt cttcggccga agttaatctc gtgtttttgg caatcaacag 120 gtttaagtcttcggccgaag ttaatctcgt gtttttggca atcaacaggt ttaagtcttc 180 ggccgaagttaatctcgtgt ttttggcaat caacaggttt aagtcttcgg ccgaagttaa 240 tctcgtgtttttggcaatca acaggtttaa gtcttcggcc gaagttaatc tcgtgttttt 300 ggcaatcaagaggtttaagt cttcggccga agttaatctc gtgtttttgg caatcaacag 360 gtttaagtcttcggccgaan ttaatctcgt gtttttggca atcaacaggt ttaantcttc 420 ggccgaagttaatctcgtgt ttttggcaat caana 455 <210> SEQ ID NO 274 <211> LENGTH: 461<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 274tttttttttt ttggccaata cccttgatga acatcaatgt gaaaatcctc ggtaaaatac 60tggcaaacca aatccagcag cacatcaaaa agcttatcca ccatgatcaa gtgggcttca 120tccctgggat gcaaggctgg ttcaacataa gaaaatcaat aaatgtaatc catcacataa 180acagaaccaa agacaaaaac cacatgatta tctcaataga tgcagaaaag gccttggaca 240aattcaacag cccttcatgc taaacactct taataaacta gatattgatg gaatgtatct 300caaaataata agagctattt atgacaaacc cacagccaat atcatactga atgggcaaag 360actggaagca ttccctttga aaactggcac aagacaagga tgccctctct caccgctcct 420attcaacata gtattggaag ttctggccag ggcaatcaag a 461 <210> SEQ ID NO 275<211> LENGTH: 729 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 164, 193, 207,215, 216, 220, 223, 241, 244, 254, 269, 271, 275, 290, 295, 298, 309,318, 325, 326, 331, 352, 380, 401, 411, 420, 424, 426, 431, 433, 435,438, 440, 442, 443, 448, 453, 464, 465, 468, 474, 475, 481, 487, 491,503, 516 <223> OTHER INFORMATION: n = A,T,C or G <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 519, 530, 531, 542, 547, 549,559, 561, 564, 582, 586, 587, 588, 589, 592, 595, 612, 614, 620, 631,632, 635, 636, 644, 646, 649, 650, 651, 655, 657, 660, 661, 662, 663,666, 672, 673, 674, 682, 687, 691, 693, 697, 700, 701, 704, 705 <223>OTHER INFORMATION: n = A,T,C or G <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 713, 715, 717, 718, 722, 726, 727 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 275 ttttttttttttggccaaca ccaagtcttc cacgtgggag gttttattat gttttacaac 60 catgaaaacataggaaggtg gctgttacag caaacatttc agatagacga atcggccaag 120 ctccccaaaccccaccttca cagcctcttc cacacgtctc ccanagattg ttgtccttca 180 cttgcaaattcanggatgtt ggaagtngac atttnnagtn gcnggaaccc catcagtgaa 240 ncantaagcagaantacgat gactttgana nacanctgat gaagaacacn ctacnganaa 300 ccctttctntcgtgttanga tctcnngtcc ntcactaatg cggccccctg cnggtccacc 360 atttgggagaactccccccn cgttggatcc ccccttgagt ntcccattct ngtcccccan 420 accngncttgngngncantn cnncctcnca ccntgtttcc ctgnngtnaa aatnngtttt 480 nccgccncccnaattcccac ccnaatcaca gcgaanccng aaggccttcn naagtgttta 540 angcccngnggtttcctcnt ntanttgcag cctaccctcc cncttnnnnt tncgngttgg 600 tcgcgccctggncncgcctn gttcctcttt nnggnnacaa cctngntcnn nggcncntcn 660 nnnctnttcctnnnactagc tngcctntcc ncnccgnggn ncanngcaca ttncncnnac 720 tntgtnncc 729<210> SEQ ID NO 276 <211> LENGTH: 339 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 276 tgacctgaca tgtagtagat acttaataaatatttgtgga atgaatggat gaagtggagt 60 tacagagaaa aatagaaaag tacaaattgttgtcagtgtt ttgaaggaaa attatgatct 120 ttcccaaagt tctgacttca ttctaagacagggttagtat ctccatacat aattttactt 180 gcttttgaaa atcaaatgag ataatctatttagattgata atttatttag actggctata 240 aactattaag tgctagcaaa tatacattttaatctcattt tccacctctt gtgatatagc 300 tatgtaggtg ttgactttaa tggatgtcaggtcaatccc 339 <210> SEQ ID NO 277 <211> LENGTH: 664 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature<222> LOCATION: 267, 534, 590, 601, 646, 657 <223> OTHER INFORMATION: n= A,T,C or G <400> SEQUENCE: 277 tgacctgaca tccataacaa aatctttctccattatattc ttctagggga atttcttgaa 60 aagcatccaa aggaaacaaa tgatggtaagaccgtgccaa gtggggagca gacaccaaag 120 taagaccaca gattttacat tcaacaggtagctcacagta ctttgcccga cactgtgggc 180 agaaatagcc tcctaatgta agccctggctcagtattgcc atccaaatgc gccatgctga 240 aagagggttt tgcatcctgg tcagatnaagaagcaatggt gtgctgagga aatcccatac 300 gaataagtga gcattcagaa cttgagctagcaggaggagg actaagatga tgtgtgagca 360 actctttgta atggctttca tctaaaataacatggtacgt gccaccagtt tcacgagcaa 420 gtacagtgca aacgcgaact tctgcagacaatccaataac agatactcta attttagctg 480 cctttagggt cttgattaaa tcataaatattagatggatc gcaagttgta aggntgctaa 540 aagatgatta gtacttctcg acttgtatgtccaggcatgt tgttttaaan tctgccttag 600 nccctgctta ggggaatttt taaagaagatggctctccat gttcanggtc aatcacnaat 660 tgcc 664 <210> SEQ ID NO 278 <211>LENGTH: 452 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 430 <223> OTHERINFORMATION: n = A,T,C or G <400> SEQUENCE: 278 tgacctgaca ttgaggaagagcacacacct ctgaaattcc ttaggttcag aagggcattt 60 gacacagagt gggcctctgataattcatga aatgcattct gaagtcatcc agaatggagg 120 ctgcaatctg ctgtgctttgggggttgcct cactgtgctc ctggatatca cacaaaagct 180 gcaatccttc ttcttcaactaacattttgc agtatttgct gggattttta ctgcagacat 240 gatacatagc ccatagtgcccagagctgaa cctctggttg agagaagttg ccaaggagcg 300 ggaaaaatgt cttgaaagatctataggtca ccaatgctgt catcttacaa cttgaacttg 360 gccaattctg tatggttgcatgcagatctt ggagaagagt acgcctctgg aagtcacggg 420 atatccaaan ctgtctgtcagatgtcaggt ca 452 <210> SEQ ID NO 279 <211> LENGTH: 274 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 279 tttttttttt ttcggcaaggcaaatttact tctgcaaaag ggtgctgctt gcacttttgg 60 ccactgcgag agcacaccaaacaaagtagg gaaggggttt ttatccctaa cgcggttatt 120 ccctggttct gtgtcgtgtccccattggct ggagtcagac tgcacaatct acactgaccc 180 aactggctac tgtttaaaattgaatatgaa taattaggta ggaaggggga ggctgtttgt 240 tacggtacaa gacgtgtttgggcatgtcag gtca 274 <210> SEQ ID NO 280 <211> LENGTH: 272 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 280 tacctgacatggagaaataa cttgtagtat tttgcgtgca atggaatact atatgagggt 60 gaaaatgaatgaactagcaa tgcgtgtatc aacatgaata aatccccaaa acataataat 120 gttgaatggaaaaggtgagt ttcagaagga tatatatgcc ctctaaatcc atttatgtaa 180 acctttaaaaaactacatta tttatggtca taagtccatc cagaaaatat ttaaaaacct 240 acatgggattgataactact gatgtcaggt ca 272 <210> SEQ ID NO 281 <211> LENGTH: 431 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 339, 420, 430, 431 <223> OTHER INFORMATION:n = A,T,C or G <400> SEQUENCE: 281 tttttttttt ttggccaata gcatgatttaaacattggaa aaagtcaaat gagcaatgcg 60 aatttttatg ttctcttgaa taatcaaaagagtaggcaac attggttcct cattcttgaa 120 tagcattaat cagaaaatat tgcatagcctctagcctcct tagagtaggt gtgctctctc 180 aaatatatca tagtcccaca gtttatttcatgtatatttt ctgcctgaat cacatagaca 240 tttgaatttg caacgcctga tgtaaatatataaattctta ccaatcagaa acatagcaag 300 aaattcaggg acttggtcat yatcagggtatgacagcana tccctgtara aacactgata 360 cacactcaca cacgtatgca acgtggagatgtcgcyttww kkktwywcwm rmrycrwcgn 420 aatcacttan n 431 <210> SEQ ID NO282 <211> LENGTH: 98 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 282 attcgattcg atgcttgagc ccaggagttc aagactgcag tgagccactgcacttcaggc 60 tggacaacag agcgagtccc tgtgccaaaa aaaaaaaa 98 <210> SEQ IDNO 283 <211> LENGTH: 764 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 372, 374,379, 380, 381, 382, 384, 387, 389, 392, 402, 409, 411, 419, 421, 432,440, 447, 452, 457, 466, 470, 471, 480, 483, 492, 503, 506, 510, 512,518, 520, 521, 524, 531, 534, 536, 542, 545, 547, 550, 552, 553, 562,566, 567, 575 <223> OTHER INFORMATION: n = A,T,C or G <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 580, 581, 584, 586, 587,595, 598, 601, 603, 604, 606, 624, 629, 630, 646, 651, 652, 653, 656,659, 664, 665, 681, 691, 700, 706, 709, 721, 724, 731, 732, 737, 741,744, 745, 750, 753, 754, 758 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 283 tttttttttt ttcgcaagca cgtgcacttt attgaatgacactgtagaca ggtgtgtggg 60 tataaactgc tgtatctagg ggcaggacca agggggcaggggcaacagcc ccagcgtgca 120 gggccascat tgcacagtgg astgcaaagg ttgcaggctatgggcggcta ctavtaaccc 180 cgtttttcct gtattatctg taacataata tggtagactgtcacagagcc gaatwccart 240 hacasgatga atccaawggt caygaggatg cccasaatcagggcccasat sttcaggcac 300 ttggcggtgg gggcatasgc ctgkgccccg gtcacgtcsccaaccwtcty cctgtcccta 360 cmcttgawtc cncnccttnn nntnccntna tntgcccgcccncctcctng ngtcaaccng 420 natctgcact anctccctcn ccccttntgg antctcntccttcaantaan nttatccttn 480 acncccccct cncctttccc ctnccncccn tnatcccngnnccnctatca ntcntnccct 540 cnctntnctn cnnatcgttc cncctnntaa ctacnctttnnacnanncct cactnatncc 600 ngnnanttct ttccttccct cccnacgcnn tgcgtgcgcccgtctngcct nnnctncgna 660 cccnnacttt atttaccttt ncaccctagc nctctacttnacccanccnc tcctacctcc 720 nggnccaccc nnccctnatc nctnnctctn tcnnctcnttcccc 764 <210> SEQ ID NO 284 <211> LENGTH: 157 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 284 caagtgtagg cacagtgatgaaagcctgga gcaaacacaa tctgtgggta attaacgttt 60 atttctcccc ttccaggaacgtcttgcatg gatgatcaaa gatcagctcc tggtcaacat 120 aaataagcta gtttaagatacgttccccta cacttga 157 <210> SEQ ID NO 285 <211> LENGTH: 150 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 285 attcgattgtactcagacaa caatatgcta agtggaagaa gtcagtcaca aaagaccaca 60 tactgtatgacttcatttac attaagtgtc cagaataggc aaatccgtag agacagaaag 120 tagatgagcagctgcctagg tctgagtaca 150 <210> SEQ ID NO 286 <211> LENGTH: 219 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 286 attcgatttttttttttttg gccatgatga aattcttact ccctcagatt ttttgtctgg 60 ataaatgcaagtctcaccac cagatgtgaa attacagtaa actttgaagg aatctcctga 120 gcaaccttggttaggatcaa tccaatattc accatctggg aagtcaggat ggctgagttg 180 caggtctttacaagttcggg ctggattggt ctgagtaca 219 <210> SEQ ID NO 287 <211> LENGTH:196 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 287attcgattct tgaggctacc aggagctagg agaagaggca tggaacaaat tttccctcat 60atccatactc agaaggaacc aaccctgctg acaccttaat ttcagcttct ggcctctaga 120actgtgagag agtacatttc tcttggttta agccaagaga atctgtcttt tggtacttta 180tatcatagcc tcaaga 196 <210> SEQ ID NO 288 <211> LENGTH: 199 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 288 attcgatttcagtccagtcc cagaacccac attgtcaatt actactctgt araagattca 60 tttgttgaaattcattgagt aaaacattta tgatccctta atatatgcca attaccatgc 120 taggtactgaagattcaagt gaccgagatg ctagcccttg ggttcaagtg atccctctcc 180 cagagtgcactggactgaa 199 <210> SEQ ID NO 289 <211> LENGTH: 182 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 289 attcgattct tgaggctacaaacctgtaca gtatgttact ctactgaata ctgtaggcaa 60 tagtaataca gaagcaagtatctgtatatg taaacattaa aaaggtacag tgaaacttca 120 gtattataat cttagggaccaccattatat atgtggtcca tcattggcca aaaaaaaaaa 180 aa 182 <210> SEQ ID NO290 <211> LENGTH: 1646 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 290 ggcacgagga gaaatgtaat tccatatttt atttgaaacttattccatat tttaattgga 60 tattgagtga ttgggttatc aaacacccac aaactttaattttgttaaat ttatatggct 120 ttgaaataga agtataagtt gctaccattt tttgataacattgaaagata gtattttacc 180 atctttaatc atcttggaaa atacaagtcc tgtgaacaaccactctttca cctagcagca 240 tgaggccaaa agtaaaggct ttaaattata acatatgggattcttagtag tatgtttttt 300 tcttgaaact cagtggctct atctaacctt actatctcctcactctttct ctaagactaa 360 actctaggct cttaaaaatc tgcccacacc aatcttagaagctctgaaaa gaatttgtct 420 ttaaatatct tttaatagta acatgtattt tatggaccaaattgacattt tcgactattt 480 tttccaaaaa agtcaggtga atttcagcac actgagttgggaatttctta tcccagaaga 540 ccaaccaatt tcatatttat ttaagattga ttccatactccgttttcaag gagaatccct 600 gcagtctcct taaaggtaga acaaatactt tctatttttttttcaccatt gtgggattgg 660 actttaagag gtgactctaa aaaaacagag aacaaatatgtctcagttgt attaagcacg 720 gacccatatt atcatattca cttaaaaaaa tgatttcctgtgcacctttt ggcaacttct 780 cttttcaatg tagggaaaaa cttagtcacc ctgaaaacccacaaaataaa taaaacttgt 840 agatgtgggc agaaggtttg ggggtggaca ttgtatgtgtttaaattaaa ccctgtatca 900 ctgagaagct gttgtatggg tcagagaaaa tgaatgcttagaagctgttc acatcttcaa 960 gagcagaagc aaaccacatg tctcagctat attattatttattttttatg cataaagtga 1020 atcatttctt ctgtattaat ttccaaaggg ttttaccctctatttaaatg ctttgaaaaa 1080 cagtgcattg acaatgggtt gatatttttc tttaaaagaaaaatataatt atgaaagcca 1140 agataatctg aagcctgttt tattttaaaa ctttttatgttctgtggttg atgttgtttg 1200 tttgtttgtt tctattttgt tggtttttta ctttgttttttgttttgttt tgttttgttt 1260 kgcatactac atgcagttct ttaaccaatg tctgtttggctaatgtaatt aaagttgtta 1320 atttatatga gtgcatttca actatgtcaa tggtttcttaatatttattg tgtagaagta 1380 ctggtaattt ttttatttac aatatgttta aagagataacagtttgatat gttttcatgt 1440 gtttatagca gaagttattt atttctatgg cattccagcggatattttgg tgtttgcgag 1500 gcatgcagtc aatattttgt acagttagtg gacagtattcagcaacgcct gatagcttct 1560 ttggccttat gttaaataaa aagacctgtt tgggatgtattttttatttt taaaaaaaaa 1620 aaaaaaaaaa aaaaaaaaaa aaaaaa 1646 <210> SEQID NO 291 <211> LENGTH: 1851 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 291 tcatcaccat tgccagcagc ggcaccgtta gtcaggttttctgggaatcc cacatgagta 60 cttccgtgtt cttcattctt cttcaatagc cataaatcttctagctctgg ctggctgttt 120 tcacttcctt taagcctttg tgactcttcc tctgatgtcagctttaagtc ttgttctgga 180 ttgctgtttt cagaagagat ttttaacatc tgtttttctttgtagtcaga aagtaactgg 240 caaattacat gatgatgact agaaacagca tactctctggccgtctttcc agatcttgag 300 aagatacatc aacattttgc tcaagtagag ggctgactatacttgctgat ccacaacata 360 cagcaagtat gagagcagtt cttccatatc tatccagcgcatttaaattc gcttttttct 420 tgattaaaaa tttcaccact tgctgttttt gctcatgtataccaagtagc agtggtgtga 480 ggccatgctt gttttttgat tcgatatcag caccgtataagagcagtgct ttggccatta 540 atttatcttc attgtagaca gcatagtgta gagtggtatttccatactca tctggaatat 600 ttggatcagt gccatgttcc agcaacatta acgcacattcatcttcctgg cattgtacgg 660 cctttgtcag agctgtcctc tttttgttgt caaggacattaagttgacat cgtctgtcca 720 gcacgagttt tactacttct gaattcccat tggcagaggccagatgtaga gcagtcctct 780 tttgcttgtc cctcttgttc acatccgtgt ccctgagcatgacgatgaga tcctttctgg 840 ggactttacc ccaccaggca gctctgtgga gcttgtccagatcttctcca tggacgtggt 900 acctgggatc catgaaggcg ctgtcatcgt agtctccccaagcgaccacg ttgctcttgc 960 cgctcccctg cagcagggga agcagtggca gcaccacttgcacctcttgc tcccaagcgt 1020 cttcacagag gagtcgttgt ggtctccaga agtgcccacgttgctcttgc cgctccccct 1080 gtccatccag ggaggaagaa atgcaggaaa tgaaagatgcatgcacgatg gtatactcct 1140 cagccatcaa acttctggac agcaggtcac ttccagcaaggtggagaaag ctgtccaccc 1200 acagaggatg agatccagaa accacaatat ccattcacaaacaaacactt ttcagccaga 1260 cacaggtact gaaatcatgt catctgcggc aacatggtggaacctaccca atcacacatc 1320 aagagatgaa gacactgcag tatatctgca caacgtaatactcttcatcc ataacaaaat 1380 aatataattt tcctctggag ccatatggat gaactatgaaggaagaactc cccgaagaag 1440 ccagtcgcag agaagccaca ctgaagctct gtcctcagccatcagcgcca cggacaggar 1500 tgtgtttctt ccccagtgat gcagcctcaa gttatcccgaagctgccgca gcacacggtg 1560 gctcctgaga aacaccccag ctcttccggt ctaacacaggcaagtcaata aatgtgataa 1620 tcacataaac agaattaaaa gcaaagtcac ataagcatctcaacagacac agaaaaggca 1680 tttgacaaaa tccagcatcc ttgtatttat tgttgcagttctcagaggaa atgcttctaa 1740 cttttcccca tttagtatta tgttggctgt gggcttgtcataggtggttt ttattacttt 1800 aaggtatgtc ccttctatgc ctgttttgct gagggttttaattctcgtgc c 1851 <210> SEQ ID NO 292 <211> LENGTH: 1851 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 292 tcatcaccat tgccagcagcggcaccgtta gtcaggtttt ctgggaatcc cacatgagta 60 cttccgtgtt cttcattcttcttcaatagc cataaatctt ctagctctgg ctggctgttt 120 tcacttcctt taagcctttgtgactcttcc tctgatgtca gctttaagtc ttgttctgga 180 ttgctgtttt cagaagagatttttaacatc tgtttttctt tgtagtcaga aagtaactgg 240 caaattacat gatgatgactagaaacagca tactctctgg ccgtctttcc agatcttgag 300 aagatacatc aacattttgctcaagtagag ggctgactat acttgctgat ccacaacata 360 cagcaagtat gagagcagttcttccatatc tatccagcgc atttaaattc gcttttttct 420 tgattaaaaa tttcaccacttgctgttttt gctcatgtat accaagtagc agtggtgtga 480 ggccatgctt gttttttgattcgatatcag caccgtataa gagcagtgct ttggccatta 540 atttatcttc attgtagacagcatagtgta gagtggtatt tccatactca tctggaatat 600 ttggatcagt gccatgttccagcaacatta acgcacattc atcttcctgg cattgtacgg 660 cctttgtcag agctgtcctctttttgttgt caaggacatt aagttgacat cgtctgtcca 720 gcacgagttt tactacttctgaattcccat tggcagaggc cagatgtaga gcagtcctct 780 tttgcttgtc cctcttgttcacatccgtgt ccctgagcat gacgatgaga tcctttctgg 840 ggactttacc ccaccaggcagctctgtgga gcttgtccag atcttctcca tggacgtggt 900 acctgggatc catgaaggcgctgtcatcgt agtctcccca agcgaccacg ttgctcttgc 960 cgctcccctg cagcaggggaagcagtggca gcaccacttg cacctcttgc tcccaagcgt 1020 cttcacagag gagtcgttgtggtctccaga agtgcccacg ttgctcttgc cgctccccct 1080 gtccatccag ggaggaagaaatgcaggaaa tgaaagatgc atgcacgatg gtatactcct 1140 cagccatcaa acttctggacagcaggtcac ttccagcaag gtggagaaag ctgtccaccc 1200 acagaggatg agatccagaaaccacaatat ccattcacaa acaaacactt ttcagccaga 1260 cacaggtact gaaatcatgtcatctgcggc aacatggtgg aacctaccca atcacacatc 1320 aagagatgaa gacactgcagtatatctgca caacgtaata ctcttcatcc ataacaaaat 1380 aatataattt tcctctggagccatatggat gaactatgaa ggaagaactc cccgaagaag 1440 ccagtcgcag agaagccacactgaagctct gtcctcagcc atcagcgcca cggacaggar 1500 tgtgtttctt ccccagtgatgcagcctcaa gttatcccga agctgccgca gcacacggtg 1560 gctcctgaga aacaccccagctcttccggt ctaacacagg caagtcaata aatgtgataa 1620 tcacataaac agaattaaaagcaaagtcac ataagcatct caacagacac agaaaaggca 1680 tttgacaaaa tccagcatccttgtatttat tgttgcagtt ctcagaggaa atgcttctaa 1740 cttttcccca tttagtattatgttggctgt gggcttgtca taggtggttt ttattacttt 1800 aaggtatgtc ccttctatgcctgttttgct gagggtttta attctcgtgc c 1851 <210> SEQ ID NO 293 <211>LENGTH: 668 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:293 cttgagcttc caaataygga agactggccc ttacacasgt caatgttaaa atgaatgcat 60ttcagtattt tgaagataaa attrgtagat ctataccttg ttttttgatt cgatatcagc 120accrtataag agcagtgctt tggccattaa tttatctttc attrtagaca gcrtagtgya 180gagtggtatt tccatactca tctggaatat ttggatcagt gccatgttcc agcaacatta 240acgcacattc atcttcctgg cattgtacgg cctgtcagta ttagacccaa aaacaaatta 300catatcttag gaattcaaaa taacattcca cagctttcac caactagtta tatttaaagg 360agaaaactca tttttatgcc atgtattgaa atcaaaccca cctcatgctg atatagttgg 420ctactgcata cctttatcag agctgtcctc tttttgttgt caaggacatt aagttgacat 480cgtctgtcca gcaggagttt tactacttct gaattcccat tggcagaggc cagatgtaga 540gcagtcctat gagagtgaga agacttttta ggaaattgta gtgcactagc tacagccata 600gcaatgattc atgtaactgc aaacactgaa tagcctgcta ttactctgcc ttcaaaaaaa 660aaaaaaaa 668 <210> SEQ ID NO 294 <211> LENGTH: 1512 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <400> SEQUENCE: 294 gggtcgccca gggggsgcgtgggctttcct cgggtgggtg tgggttttcc ctgggtgggg 60 tgggctgggc trgaatcccctgctggggtt ggcaggtttt ggctgggatt gacttttytc 120 ttcaaacaga ttggaaacccggagttacct gctagttggt gaaactggtt ggtagacgcg 180 atctgttggc tactactggcttctcctggc tgttaaaagc agatggtggt tgaggttgat 240 tccatgccgg ctgcttcttctgtgaagaag ccatttggtc tcaggagcaa gatgggcaag 300 tggtgctgcc gttgcttcccctgctgcagg gagagcggca agagcaacgt gggcacttct 360 ggagaccacg acgactctgctatgaagaca ctcaggagca agatgggcaa gtggtgccgc 420 cactgcttcc cctgctgcagggggagtggc aagagcaacg tgggcgcttc tggagaccac 480 gacgaytctg ctatgaagacactcaggaac aagatgggca agtggtgctg ccactgcttc 540 ccctgctgca gggggagcrgcaagagcaag gtgggcgctt ggggagacta cgatgacagt 600 gccttcatgg agcccaggtaccacgtccgt ggagaagatc tggacaagct ccacagagct 660 gcctggtggg gtaaagtccccagaaaggat ctcatcgtca tgctcaggga cactgacgtg 720 aacaagaagg acaagcaaaagaggactgct ctacatctgg cctctgccaa tgggaattca 780 gaagtagtaa aactcstgctggacagacga tgtcaactta atgtccttga caacaaaaag 840 aggacagctc tgayaaaggccgtacaatgc caggaagatg aatgtgcgtt aatgttgctg 900 gaacatggca ctgatccaaatattccagat gagtatggaa ataccactct rcactaygct 960 rtctayaatg aagataaattaatggccaaa gcactgctct tatayggtgc tgatatcgaa 1020 tcaaaaaaca aggtatagatctactaattt tatcttcaaa atactgaaat gcattcattt 1080 taacattgac gtgtgtaagggccagtcttc cgtatttgga agctcaagca taacttgaat 1140 gaaaatattt tgaaatgacctaattatctm agactttatt ttaaatattg ttattttcaa 1200 agaagcatta gagggtacagtttttttttt ttaaatgcac ttctggtaaa tacttttgtt 1260 gaaaacactg aatttgtaaaaggtaatact tactattttt caatttttcc ctcctaggat 1320 ttttttcccc taatgaatgtaagatggcaa aatttgccct gaaataggtt ttacatgaaa 1380 actccaagaa aagttaaacatgtttcagtg aatagagatc ctgctccttt ggcaagttcc 1440 taaaaaacag taatagatacgaggtgatgc gcctgtcagt ggcaaggttt aagatatttc 1500 tgatctcgtg cc 1512<210> SEQ ID NO 295 <211> LENGTH: 1853 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 295 gggtcgccca gggggsgcgt gggctttcctcgggtgggtg tgggttttcc ctgggtgggg 60 tgggctgggc trgaatcccc tgctggggttggcaggtttt ggctgggatt gacttttytc 120 ttcaaacaga ttggaaaccc ggagttacctgctagttggt gaaactggtt ggtagacgcg 180 atctgttggc tactactggc ttctcctggctgttaaaagc agatggtggt tgaggttgat 240 tccatgccgg ctgcttcttc tgtgaagaagccatttggtc tcaggagcaa gatgggcaag 300 tggtgctgcc gttgcttccc ctgctgcagggagagcggca agagcaacgt gggcacttct 360 ggagaccacg acgactctgc tatgaagacactcaggagca agatgggcaa gtggtgccgc 420 cactgcttcc cctgctgcag ggggagtggcaagagcaacg tgggcgcttc tggagaccac 480 gacgaytctg ctatgaagac actcaggaacaagatgggca agtggtgctg ccactgcttc 540 ccctgctgca gggggagcrg caagagcaaggtgggcgctt ggggagacta cgatgacagy 600 gccttcatgg akcccaggta ccacgtccrtggagaagatc tggacaagct ccacagagct 660 gcctggtggg gtaaagtccc cagaaaggatctcatcgtca tgctcaggga cackgaygtg 720 aacaagargg acaagcaaaa gaggactgctctacatctgg cctctgccaa tgggaattca 780 gaagtagtaa aactcstgct ggacagacgatgtcaactta atgtccttga caacaaaaag 840 aggacagctc tgayaaaggc cgtacaatgccaggaagatg aatgtgcgtt aatgttgctg 900 gaacatggca ctgatccaaa tattccagatgagtatggaa ataccactct rcactaygct 960 rtctayaatg aagataaatt aatggccaaagcactgctct tatayggtgc tgatatcgaa 1020 tcaaaaaaca agcatggcct cacaccactgytacttggtr tacatgagca aaaacagcaa 1080 gtsgtgaaat ttttaatyaa gaaaaaagcgaatttaaaat gcrctggata gatatggaag 1140 ractgctctc atacttgctg tatgttgtggatcagcaagt atagtcagcc ytctacttga 1200 gcaaaatrtt gatgtatctt ctcaagatctggaaagacgg ccagagagta tgctgtttct 1260 agtcatcatc atgtaatttg ccagttactttctgactaca aagaaaaaca gatgttaaaa 1320 atctcttctg aaaacagcaa tccagaacaagacttaaagc tgacatcaga ggaagagtca 1380 caaaggctta aaggaagtga aaacagccagccagaggcat ggaaactttt aaatttaaac 1440 ttttggttta atgttttttt tttttgccttaataatatta gatagtccca aatgaaatwa 1500 cctatgagac taggctttga gaatcaatagattctttttt taagaatctt ttggctagga 1560 gcggtgtctc acgcctgtaa ttccagcaccttgagaggct gaggtgggca gatcacgaga 1620 tcaggagatc gagaccatcc tggctaacacggtgaaaccc catctctact aaaaatacaa 1680 aaacttagct gggtgtggtg gcgggtgcctgtagtcccag ctactcagga rgctgaggca 1740 ggagaatggc atgaacccgg gaggtggaggttgcagtgag ccgagatccg ccactacact 1800 ccagcctggg tgacagagca agactctgtctcaaaaaaaa aaaaaaaaaa aaa 1853 <210> SEQ ID NO 296 <211> LENGTH: 2184<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 296ggcacgagaa ttaaaaccct cagcaaaaca ggcatagaag ggacatacct taaagtaata 60aaaaccacct atgacaagcc cacagccaac ataatactaa atggggaaaa gttagaagca 120tttcctctga gaactgcaac aataaataca aggatgctgg attttgtcaa atgccttttc 180tgtgtctgtt gagatgctta tgtgactttg cttttaattc tgtttatgtg attatcacat 240ttattgactt gcctgtgtta gaccggaaga gctggggtgt ttctcaggag ccaccgtgtg 300ctgcggcagc ttcgggataa cttgaggctg catcactggg gaagaaacac aytcctgtcc 360gtggcgctga tggctgagga cagagcttca gtgtggcttc tctgcgactg gcttcttcgg 420ggagttcttc cttcatagtt catccatatg gctccagagg aaaattatat tattttgtta 480tggatgaaga gtattacgtt gtgcagatat actgcagtgt cttcatctct tgatgtgtga 540ttgggtaggt tccaccatgt tgccgcagat gacatgattt cagtacctgt gtctggctga 600aaagtgtttg tttgtgaatg gatattgtgg tttctggatc tcatcctctg tgggtggaca 660gctttctcca ccttgctgga agtgacctgc tgtccagaag tttgatggct gaggagtata 720ccatcgtgca tgcatctttc atttcctgca tttcttcctc cctggatgga cagggggagc 780ggcaagagca acgtgggcac ttctggagac cacaacgact cctctgtgaa gacgcttggg 840agcaagaggt gcaagtggtg ctgccactgc ttcccctgct gcaggggagc ggcaagagca 900acgtggtcgc ttggggagac tacgatgaca gcgccttcat ggatcccagg taccacgtcc 960atggagaaga tctggacaag ctccacagag ctgcctggtg gggtaaagtc cccagaaagg 1020atctcatcgt catgctcagg gacacggatg tgaacaagag ggacaagcaa aagaggactg 1080ctctacatct ggcctctgcc aatgggaatt cagaagtagt aaaactcgtg ctggacagac 1140gatgtcaact taatgtcctt gacaacaaaa agaggacagc tctgacaaag gccgtacaat 1200gccaggaaga tgaatgtgcg ttaatgttgc tggaacatgg cactgatcca aatattccag 1260atgagtatgg aaataccact ctacactatg ctgtctacaa tgaagataaa ttaatggcca 1320aagcactgct cttatacggt gctgatatcg aatcaaaaaa caagcatggc ctcacaccac 1380tgctacttgg tatacatgag caaaaacagc aagtggtgaa atttttaatc aagaaaaaag 1440cgaatttaaa tgcgctggat agatatggaa gaactgctct catacttgct gtatgttgtg 1500gatcagcaag tatagtcagc cctctacttg agcaaaatgt tgatgtatct tctcaagatc 1560tggaaagacg gccagagagt atgctgtttc tagtcatcat catgtaattt gccagttact 1620ttctgactac aaagaaaaac agatgttaaa aatctcttct gaaaacagca atccagaaca 1680agacttaaag ctgacatcag aggaagagtc acaaaggctt aaaggaagtg aaaacagcca 1740gccagaggca tggaaacttt taaatttaaa cttttggttt aatgtttttt ttttttgcct 1800taataatatt agatagtccc aaatgaaatw acctatgaga ctaggctttg agaatcaata 1860gattcttttt ttaagaatct tttggctagg agcggtgtct cacgcctgta attccagcac 1920cttgagaggc tgaggtgggc agatcacgag atcaggagat cgagaccatc ctggctaaca 1980cggtgaaacc ccatctctac taaaaataca aaaacttagc tgggtgtggt ggcgggtgcc 2040tgtagtccca gctactcagg argctgaggc aggagaatgg catgaacccg ggaggtggag 2100gttgcagtga gccgagatcc gccactacac tccagcctgg gtgacagagc aagactctgt 2160ctcaaaaaaa aaaaaaaaaa aaaa 2184 <210> SEQ ID NO 297 <211> LENGTH: 1855<212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 606 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 297 tgcacgcatc ggccagtgtc tgtgccacgtacactgacgc cccctgagat gtgcacgccg 60 cacgcgcacg ttgcacgcgc ggcagcggcttggctggctt gtaacggctt gcacgcgcac 120 gccgcccccg cataaccgtc agactggcctgtaacggctt gcaggcgcac gccgcacgcg 180 cgtaacggct tggctgccct gtaacggcttgcacgtgcat gctgcacgcg cgttaacggc 240 ttggctggca tgtagccgct tggcttggctttgcattytt tgctkggctk ggcgttgkty 300 tcttggattg acgcttcctc cttggatkgacgtttcctcc ttggatkgac gtttcytyty 360 tcgcgttcct ttgctggact tgaccttttytctgctgggt ttggcattcc tttggggtgg 420 gctgggtgtt ttctccgggg gggktkgcccttcctggggt gggcgtgggk cgcccccagg 480 gggcgtgggc tttccccggg tgggtgtgggttttcctggg gtggggtggg ctgtgctggg 540 atccccctgc tggggttggc agggattgacttttttcttc aaacagattg gaaacccgga 600 gtaacntgct agttggtgaa actggttggtagacgcgatc tgctggtact actgtttctc 660 ctggctgtta aaagcagatg gtggctgaggttgattcaat gccggctgct tcttctgtga 720 agaagccatt tggtctcagg agcaagatgggcaagtggtg cgccactgct tcccctgctg 780 cagggggagc ggcaagagca acgtgggcacttctggagac cacaacgact cctctgtgaa 840 gacgcttggg agcaagaggt gcaagtggtgctgcccactg cttcccctgc tgcaggggag 900 cggcaagagc aacgtggkcg cttggggagactacgatgac agcgccttca tggakcccag 960 gtaccacgtc crtggagaag atctggacaagctccacaga gctgcctggt ggggtaaagt 1020 ccccagaaag gatctcatcg tcatgctcagggacactgay gtgaacaaga rggacaagca 1080 aaagaggact gctctacatc tggcctctgccaatgggaat tcagaagtag taaaactcgt 1140 gctggacaga cgatgtcaac ttaatgtccttgacaacaaa aagaggacag ctctgacaaa 1200 ggccgtacaa tgccaggaag atgaatgtgcgttaatgttg ctggaacatg gcactgatcc 1260 aaatattcca gatgagtatg gaaataccactctacactat gctgtctaca atgaagataa 1320 attaatggcc aaagcactgc tcttatacggtgctgatatc gaatcaaaaa acaaggtata 1380 gatctactaa ttttatcttc aaaatactgaaatgcattca ttttaacatt gacgtgtgta 1440 agggccagtc ttccgtattt ggaagctcaagcataacttg aatgaaaata ttttgaaatg 1500 acctaattat ctaagacttt attttaaatattgttatttt caaagaagca ttagagggta 1560 cagttttttt tttttaaatg cacttctggtaaatactttt gttgaaaaca ctgaatttgt 1620 aaaaggtaat acttactatt tttcaatttttccctcctag gatttttttc ccctaatgaa 1680 tgtaagatgg caaaatttgc cctgaaataggttttacatg aaaactccaa gaaaagttaa 1740 acatgtttca gtgaatagag atcctgctcctttggcaagt tcctaaaaaa cagtaataga 1800 tacgaggtga tgcgcctgtc agtggcaaggtttaagatat ttctgatctc gtgcc 1855 <210> SEQ ID NO 298 <211> LENGTH: 1059<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 298gcaacgtggg cacttctgga gaccacaacg actcctctgt gaagacgctt gggagcaaga 60ggtgcaagtg gtgctgccca ctgcttcccc tgctgcaggg gagcggcaag agcaacgtgg 120gcgcttgrgg agactmcgat gacagygcct tcatggagcc caggtaccac gtccgtggag 180aagatctgga caagctccac agagctgccc tggtggggta aagtccccag aaaggatctc 240atcgtcatgc tcagggacac tgaygtgaac aagarggaca agcaaaagag gactgctcta 300catctggcct ctgccaatgg gaattcagaa gtagtaaaac tcstgctgga cagacgatgt 360caacttaatg tccttgacaa caaaaagagg acagctctga yaaaggccgt acaatgccag 420gaagatgaat gtgcgttaat gttgctggaa catggcactg atccaaatat tccagatgag 480tatggaaata ccactctrca ctaygctrtc tayaatgaag ataaattaat ggccaaagca 540ctgctcttat ayggtgctga tatcgaatca aaaaacaagg tatagatcta ctaattttat 600cttcaaaata ctgaaatgca ttcattttaa cattgacgtg tgtaagggcc agtcttccgt 660atttggaagc tcaagcataa cttgaatgaa aatattttga aatgacctaa ttatctaaga 720ctttatttta aatattgtta ttttcaaaga agcattagag ggtacagttt ttttttttta 780aatgcacttc tggtaaatac ttttgttgaa aacactgaat ttgtaaaagg taatacttac 840tatttttcaa tttttccctc ctaggatttt tttcccctaa tgaatgtaag atggcaaaat 900ttgccctgaa ataggtttta catgaaaact ccaagaaaag ttaaacatgt ttcagtgaat 960agagatcctg ctcctttggc aagttcctaa aaaacagtaa tagatacgag gtgatgcgcc 1020tgtcagtggc aaggtttaag atatttctga tctcgtgcc 1059 <210> SEQ ID NO 299<211> LENGTH: 329 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 299 Met Asp Ile Val Val Ser Gly Ser His Pro Leu Trp Val AspSer Phe 1 5 10 15 Leu His Leu Ala Gly Ser Asp Leu Leu Ser Arg Ser LeuMet Ala Glu 20 25 30 Glu Tyr Thr Ile Val His Ala Ser Phe Ile Ser Cys IleSer Ser Ser 35 40 45 Leu Asp Gly Gln Gly Glu Arg Gln Glu Gln Arg Gly HisPhe Trp Arg 50 55 60 Pro Gln Arg Leu Leu Cys Glu Asp Ala Trp Glu Gln GluVal Gln Val 65 70 75 80 Val Leu Pro Leu Leu Pro Leu Leu Gln Gly Ser GlyLys Ser Asn Val 85 90 95 Val Ala Trp Gly Asp Tyr Asp Asp Ser Ala Phe MetAsp Pro Arg Tyr 100 105 110 His Val His Gly Glu Asp Leu Asp Lys Leu HisArg Ala Ala Trp Trp 115 120 125 Gly Lys Val Pro Arg Lys Asp Leu Ile ValMet Leu Arg Asp Thr Asp 130 135 140 Val Asn Lys Arg Asp Lys Gln Lys ArgThr Ala Leu His Leu Ala Ser 145 150 155 160 Ala Asn Gly Asn Ser Glu ValVal Lys Leu Val Leu Asp Arg Arg Cys 165 170 175 Gln Leu Asn Val Leu AspAsn Lys Lys Arg Thr Ala Leu Thr Lys Ala 180 185 190 Val Gln Cys Gln GluAsp Glu Cys Ala Leu Met Leu Leu Glu His Gly 195 200 205 Thr Asp Pro AsnIle Pro Asp Glu Tyr Gly Asn Thr Thr Leu His Tyr 210 215 220 Ala Val TyrAsn Glu Asp Lys Leu Met Ala Lys Ala Leu Leu Leu Tyr 225 230 235 240 GlyAla Asp Ile Glu Ser Lys Asn Lys His Gly Leu Thr Pro Leu Leu 245 250 255Leu Gly Ile His Glu Gln Lys Gln Gln Val Val Lys Phe Leu Ile Lys 260 265270 Lys Lys Ala Asn Leu Asn Ala Leu Asp Arg Tyr Gly Arg Thr Ala Leu 275280 285 Ile Leu Ala Val Cys Cys Gly Ser Ala Ser Ile Val Ser Pro Leu Leu290 295 300 Glu Gln Asn Val Asp Val Ser Ser Gln Asp Leu Glu Arg Arg ProGlu 305 310 315 320 Ser Met Leu Phe Leu Val Ile Ile Met 325 <210> SEQ IDNO 300 <211> LENGTH: 148 <212> TYPE: PRT <213> ORGANISM: Homo sapiens<220> FEATURE: <221> NAME/KEY: VARIANT <222> LOCATION: 3, 46, 69, 88,124 <223> OTHER INFORMATION: Xaa = Any Amino Acid <400> SEQUENCE: 300Met Thr Xaa Pro Ser Trp Ser Pro Gly Thr Thr Ser Val Glu Lys Ile 1 5 1015 Trp Thr Ser Ser Thr Glu Leu Pro Trp Trp Gly Lys Val Pro Arg Lys 20 2530 Asp Leu Ile Val Met Leu Arg Asp Thr Asp Val Asn Lys Xaa Asp Lys 35 4045 Gln Lys Arg Thr Ala Leu His Leu Ala Ser Ala Asn Gly Asn Ser Glu 50 5560 Val Val Lys Leu Xaa Leu Asp Arg Arg Cys Gln Leu Asn Val Leu Asp 65 7075 80 Asn Lys Lys Arg Thr Ala Leu Xaa Lys Ala Val Gln Cys Gln Glu Asp 8590 95 Glu Cys Ala Leu Met Leu Leu Glu His Gly Thr Asp Pro Asn Ile Pro100 105 110 Asp Glu Tyr Gly Asn Thr Thr Leu His Tyr Ala Xaa Tyr Asn GluAsp 115 120 125 Lys Leu Met Ala Lys Ala Leu Leu Leu Tyr Gly Ala Asp IleGlu Ser 130 135 140 Lys Asn Lys Val 145 <210> SEQ ID NO 301 <211>LENGTH: 1155 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 301 atggtggttg aggttgattc catgccggct gcctcttctg tgaagaagccatttggtctc 60 aggagcaaga tgggcaagtg gtgctgccgt tgcttcccct gctgcagggagagcggcaag 120 agcaacgtgg gcacttctgg agaccacgac gactctgcta tgaagacactcaggagcaag 180 atgggcaagt ggtgccgcca ctgcttcccc tgctgcaggg ggagtggcaagagcaacgtg 240 ggcgcttctg gagaccacga cgactctgct atgaagacac tcaggaacaagatgggcaag 300 tggtgctgcc actgcttccc ctgctgcagg gggagcggca agagcaaggtgggcgcttgg 360 ggagactacg atgacagtgc cttcatggag cccaggtacc acgtccgtggagaagatctg 420 gacaagctcc acagagctgc ctggtggggt aaagtcccca gaaaggatctcatcgtcatg 480 ctcagggaca ctgacgtgaa caagaaggac aagcaaaaga ggactgctctacatctggcc 540 tctgccaatg ggaattcaga agtagtaaaa ctcctgctgg acagacgatgtcaacttaat 600 gtccttgaca acaaaaagag gacagctctg ataaaggccg tacaatgccaggaagatgaa 660 tgtgcgttaa tgttgctgga acatggcact gatccaaata ttccagatgagtatggaaat 720 accactctgc actacgctat ctataatgaa gataaattaa tggccaaagcactgctctta 780 tatggtgctg atatcgaatc aaaaaacaag catggcctca caccactgttacttggtgta 840 catgagcaaa aacagcaagt cgtgaaattt ttaatcaaga aaaaagcgaatttaaatgca 900 ctggatagat atggaaggac tgctctcata cttgctgtat gttgtggatcagcaagtata 960 gtcagccttc tacttgagca aaatattgat gtatcttctc aagatctatctggacagacg 1020 gccagagagt atgctgtttc tagtcatcat catgtaattt gccagttactttctgactac 1080 aaagaaaaac agatgctaaa aatctcttct gaaaacagca atccagaaaatgtctcaaga 1140 accagaaata aataa 1155 <210> SEQ ID NO 302 <211> LENGTH:2000 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 302atggtggttg aggttgattc catgccggct gcctcttctg tgaagaagcc atttggtctc 60aggagcaaga tgggcaagtg gtgctgccgt tgcttcccct gctgcaggga gagcggcaag 120agcaacgtgg gcacttctgg agaccacgac gactctgcta tgaagacact caggagcaag 180atgggcaagt ggtgccgcca ctgcttcccc tgctgcaggg ggagtggcaa gagcaacgtg 240ggcgcttctg gagaccacga cgactctgct atgaagacac tcaggaacaa gatgggcaag 300tggtgctgcc actgcttccc ctgctgcagg gggagcggca agagcaaggt gggcgcttgg 360ggagactacg atgacagtgc cttcatggag cccaggtacc acgtccgtgg agaagatctg 420gacaagctcc acagagctgc ctggtggggt aaagtcccca gaaaggatct catcgtcatg 480ctcagggaca ctgacgtgaa caagaaggac aagcaaaaga ggactgctct acatctggcc 540tctgccaatg ggaattcaga agtagtaaaa ctcctgctgg acagacgatg tcaacttaat 600gtccttgaca acaaaaagag gacagctctg ataaaggccg tacaatgcca ggaagatgaa 660tgtgcgttaa tgttgctgga acatggcact gatccaaata ttccagatga gtatggaaat 720accactctgc actacgctat ctataatgaa gataaattaa tggccaaagc actgctctta 780tatggtgctg atatcgaatc aaaaaacaag catggcctca caccactgtt acttggtgta 840catgagcaaa aacagcaagt cgtgaaattt ttaatcaaga aaaaagcgaa tttaaatgca 900ctggatagat atggaaggac tgctctcata cttgctgtat gttgtggatc agcaagtata 960gtcagccttc tacttgagca aaatattgat gtatcttctc aagatctatc tggacagacg 1020gccagagagt atgctgtttc tagtcatcat catgtaattt gccagttact ttctgactac 1080aaagaaaaac agatgctaaa aatctcttct gaaaacagca atccagaaca agacttaaag 1140ctgacatcag aggaagagtc acaaaggttc aaaggcagtg aaaatagcca gccagagaaa 1200atgtctcaag aaccagaaat aaataaggat ggtgatagag aggttgaaga agaaatgaag 1260aagcatgaaa gtaataatgt gggattacta gaaaacctga ctaatggtgt cactgctggc 1320aatggtgata atggattaat tcctcaaagg aagagcagaa cacctgaaaa tcagcaattt 1380cctgacaacg aaagtgaaga gtatcacaga atttgcgaat tagtttctga ctacaaagaa 1440aaacagatgc caaaatactc ttctgaaaac agcaacccag aacaagactt aaagctgaca 1500tcagaggaag agtcacaaag gcttgagggc agtgaaaatg gccagccaga gctagaaaat 1560tttatggcta tcgaagaaat gaagaagcac ggaagtactc atgtcggatt cccagaaaac 1620ctgactaatg gtgccactgc tggcaatggt gatgatggat taattcctcc aaggaagagc 1680agaacacctg aaagccagca atttcctgac actgagaatg aagagtatca cagtgacgaa 1740caaaatgata ctcagaagca attttgtgaa gaacagaaca ctggaatatt acacgatgag 1800attctgattc atgaagaaaa gcagatagaa gtggttgaaa aaatgaattc tgagctttct 1860cttagttgta agaaagaaaa agacatcttg catgaaaata gtacgttgcg ggaagaaatt 1920gccatgctaa gactggagct agacacaatg aaacatcaga gccagctaaa aaaaaaaaaa 1980aaaaaaaaaa aaaaaaaaaa 2000 <210> SEQ ID NO 303 <211> LENGTH: 2040 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 303 atggtggttgaggttgattc catgccggct gcctcttctg tgaagaagcc atttggtctc 60 aggagcaagatgggcaagtg gtgctgccgt tgcttcccct gctgcaggga gagcggcaag 120 agcaacgtgggcacttctgg agaccacgac gactctgcta tgaagacact caggagcaag 180 atgggcaagtggtgccgcca ctgcttcccc tgctgcaggg ggagtggcaa gagcaacgtg 240 ggcgcttctggagaccacga cgactctgct atgaagacac tcaggaacaa gatgggcaag 300 tggtgctgccactgcttccc ctgctgcagg gggagcggca agagcaaggt gggcgcttgg 360 ggagactacgatgacagtgc cttcatggag cccaggtacc acgtccgtgg agaagatctg 420 gacaagctccacagagctgc ctggtggggt aaagtcccca gaaaggatct catcgtcatg 480 ctcagggacactgacgtgaa caagaaggac aagcaaaaga ggactgctct acatctggcc 540 tctgccaatgggaattcaga agtagtaaaa ctcctgctgg acagacgatg tcaacttaat 600 gtccttgacaacaaaaagag gacagctctg ataaaggccg tacaatgcca ggaagatgaa 660 tgtgcgttaatgttgctgga acatggcact gatccaaata ttccagatga gtatggaaat 720 accactctgcactacgctat ctataatgaa gataaattaa tggccaaagc actgctctta 780 tatggtgctgatatcgaatc aaaaaacaag catggcctca caccactgtt acttggtgta 840 catgagcaaaaacagcaagt cgtgaaattt ttaatcaaga aaaaagcgaa tttaaatgca 900 ctggatagatatggaaggac tgctctcata cttgctgtat gttgtggatc agcaagtata 960 gtcagccttctacttgagca aaatattgat gtatcttctc aagatctatc tggacagacg 1020 gccagagagtatgctgtttc tagtcatcat catgtaattt gccagttact ttctgactac 1080 aaagaaaaacagatgctaaa aatctcttct gaaaacagca atccagaaca agacttaaag 1140 ctgacatcagaggaagagtc acaaaggttc aaaggcagtg aaaatagcca gccagagaaa 1200 atgtctcaagaaccagaaat aaataaggat ggtgatagag aggttgaaga agaaatgaag 1260 aagcatgaaagtaataatgt gggattacta gaaaacctga ctaatggtgt cactgctggc 1320 aatggtgataatggattaat tcctcaaagg aagagcagaa cacctgaaaa tcagcaattt 1380 cctgacaacgaaagtgaaga gtatcacaga atttgcgaat tagtttctga ctacaaagaa 1440 aaacagatgccaaaatactc ttctgaaaac agcaacccag aacaagactt aaagctgaca 1500 tcagaggaagagtcacaaag gcttgagggc agtgaaaatg gccagccaga gaaaagatct 1560 caagaaccagaaataaataa ggatggtgat agagagctag aaaattttat ggctatcgaa 1620 gaaatgaagaagcacggaag tactcatgtc ggattcccag aaaacctgac taatggtgcc 1680 actgctggcaatggtgatga tggattaatt cctccaagga agagcagaac acctgaaagc 1740 cagcaatttcctgacactga gaatgaagag tatcacagtg acgaacaaaa tgatactcag 1800 aagcaattttgtgaagaaca gaacactgga atattacacg atgagattct gattcatgaa 1860 gaaaagcagatagaagtggt tgaaaaaatg aattctgagc tttctcttag ttgtaagaaa 1920 gaaaaagacatcttgcatga aaatagtacg ttgcgggaag aaattgccat gctaagactg 1980 gagctagacacaatgaaaca tcagagccag ctaaaaaaaa aaaaaaaaaa aaaaaaaaaa 2040 <210> SEQ IDNO 304 <211> LENGTH: 384 <212> TYPE: PRT <213> ORGANISM: Homo sapiens<400> SEQUENCE: 304 Met Val Val Glu Val Asp Ser Met Pro Ala Ala Ser SerVal Lys Lys 1 5 10 15 Pro Phe Gly Leu Arg Ser Lys Met Gly Lys Trp CysCys Arg Cys Phe 20 25 30 Pro Cys Cys Arg Glu Ser Gly Lys Ser Asn Val GlyThr Ser Gly Asp 35 40 45 His Asp Asp Ser Ala Met Lys Thr Leu Arg Ser LysMet Gly Lys Trp 50 55 60 Cys Arg His Cys Phe Pro Cys Cys Arg Gly Ser GlyLys Ser Asn Val 65 70 75 80 Gly Ala Ser Gly Asp His Asp Asp Ser Ala MetLys Thr Leu Arg Asn 85 90 95 Lys Met Gly Lys Trp Cys Cys His Cys Phe ProCys Cys Arg Gly Ser 100 105 110 Gly Lys Ser Lys Val Gly Ala Trp Gly AspTyr Asp Asp Ser Ala Phe 115 120 125 Met Glu Pro Arg Tyr His Val Arg GlyGlu Asp Leu Asp Lys Leu His 130 135 140 Arg Ala Ala Trp Trp Gly Lys ValPro Arg Lys Asp Leu Ile Val Met 145 150 155 160 Leu Arg Asp Thr Asp ValAsn Lys Lys Asp Lys Gln Lys Arg Thr Ala 165 170 175 Leu His Leu Ala SerAla Asn Gly Asn Ser Glu Val Val Lys Leu Leu 180 185 190 Leu Asp Arg ArgCys Gln Leu Asn Val Leu Asp Asn Lys Lys Arg Thr 195 200 205 Ala Leu IleLys Ala Val Gln Cys Gln Glu Asp Glu Cys Ala Leu Met 210 215 220 Leu LeuGlu His Gly Thr Asp Pro Asn Ile Pro Asp Glu Tyr Gly Asn 225 230 235 240Thr Thr Leu His Tyr Ala Ile Tyr Asn Glu Asp Lys Leu Met Ala Lys 245 250255 Ala Leu Leu Leu Tyr Gly Ala Asp Ile Glu Ser Lys Asn Lys His Gly 260265 270 Leu Thr Pro Leu Leu Leu Gly Val His Glu Gln Lys Gln Gln Val Val275 280 285 Lys Phe Leu Ile Lys Lys Lys Ala Asn Leu Asn Ala Leu Asp ArgTyr 290 295 300 Gly Arg Thr Ala Leu Ile Leu Ala Val Cys Cys Gly Ser AlaSer Ile 305 310 315 320 Val Ser Leu Leu Leu Glu Gln Asn Ile Asp Val SerSer Gln Asp Leu 325 330 335 Ser Gly Gln Thr Ala Arg Glu Tyr Ala Val SerSer His His His Val 340 345 350 Ile Cys Gln Leu Leu Ser Asp Tyr Lys GluLys Gln Met Leu Lys Ile 355 360 365 Ser Ser Glu Asn Ser Asn Pro Glu AsnVal Ser Arg Thr Arg Asn Lys 370 375 380 <210> SEQ ID NO 305 <211>LENGTH: 656 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:305 Met Val Val Glu Val Asp Ser Met Pro Ala Ala Ser Ser Val Lys Lys 1 510 15 Pro Phe Gly Leu Arg Ser Lys Met Gly Lys Trp Cys Cys Arg Cys Phe 2025 30 Pro Cys Cys Arg Glu Ser Gly Lys Ser Asn Val Gly Thr Ser Gly Asp 3540 45 His Asp Asp Ser Ala Met Lys Thr Leu Arg Ser Lys Met Gly Lys Trp 5055 60 Cys Arg His Cys Phe Pro Cys Cys Arg Gly Ser Gly Lys Ser Asn Val 6570 75 80 Gly Ala Ser Gly Asp His Asp Asp Ser Ala Met Lys Thr Leu Arg Asn85 90 95 Lys Met Gly Lys Trp Cys Cys His Cys Phe Pro Cys Cys Arg Gly Ser100 105 110 Gly Lys Ser Lys Val Gly Ala Trp Gly Asp Tyr Asp Asp Ser AlaPhe 115 120 125 Met Glu Pro Arg Tyr His Val Arg Gly Glu Asp Leu Asp LysLeu His 130 135 140 Arg Ala Ala Trp Trp Gly Lys Val Pro Arg Lys Asp LeuIle Val Met 145 150 155 160 Leu Arg Asp Thr Asp Val Asn Lys Lys Asp LysGln Lys Arg Thr Ala 165 170 175 Leu His Leu Ala Ser Ala Asn Gly Asn SerGlu Val Val Lys Leu Leu 180 185 190 Leu Asp Arg Arg Cys Gln Leu Asn ValLeu Asp Asn Lys Lys Arg Thr 195 200 205 Ala Leu Ile Lys Ala Val Gln CysGln Glu Asp Glu Cys Ala Leu Met 210 215 220 Leu Leu Glu His Gly Thr AspPro Asn Ile Pro Asp Glu Tyr Gly Asn 225 230 235 240 Thr Thr Leu His TyrAla Ile Tyr Asn Glu Asp Lys Leu Met Ala Lys 245 250 255 Ala Leu Leu LeuTyr Gly Ala Asp Ile Glu Ser Lys Asn Lys His Gly 260 265 270 Leu Thr ProLeu Leu Leu Gly Val His Glu Gln Lys Gln Gln Val Val 275 280 285 Lys PheLeu Ile Lys Lys Lys Ala Asn Leu Asn Ala Leu Asp Arg Tyr 290 295 300 GlyArg Thr Ala Leu Ile Leu Ala Val Cys Cys Gly Ser Ala Ser Ile 305 310 315320 Val Ser Leu Leu Leu Glu Gln Asn Ile Asp Val Ser Ser Gln Asp Leu 325330 335 Ser Gly Gln Thr Ala Arg Glu Tyr Ala Val Ser Ser His His His Val340 345 350 Ile Cys Gln Leu Leu Ser Asp Tyr Lys Glu Lys Gln Met Leu LysIle 355 360 365 Ser Ser Glu Asn Ser Asn Pro Glu Gln Asp Leu Lys Leu ThrSer Glu 370 375 380 Glu Glu Ser Gln Arg Phe Lys Gly Ser Glu Asn Ser GlnPro Glu Lys 385 390 395 400 Met Ser Gln Glu Pro Glu Ile Asn Lys Asp GlyAsp Arg Glu Val Glu 405 410 415 Glu Glu Met Lys Lys His Glu Ser Asn AsnVal Gly Leu Leu Glu Asn 420 425 430 Leu Thr Asn Gly Val Thr Ala Gly AsnGly Asp Asn Gly Leu Ile Pro 435 440 445 Gln Arg Lys Ser Arg Thr Pro GluAsn Gln Gln Phe Pro Asp Asn Glu 450 455 460 Ser Glu Glu Tyr His Arg IleCys Glu Leu Val Ser Asp Tyr Lys Glu 465 470 475 480 Lys Gln Met Pro LysTyr Ser Ser Glu Asn Ser Asn Pro Glu Gln Asp 485 490 495 Leu Lys Leu ThrSer Glu Glu Glu Ser Gln Arg Leu Glu Gly Ser Glu 500 505 510 Asn Gly GlnPro Glu Leu Glu Asn Phe Met Ala Ile Glu Glu Met Lys 515 520 525 Lys HisGly Ser Thr His Val Gly Phe Pro Glu Asn Leu Thr Asn Gly 530 535 540 AlaThr Ala Gly Asn Gly Asp Asp Gly Leu Ile Pro Pro Arg Lys Ser 545 550 555560 Arg Thr Pro Glu Ser Gln Gln Phe Pro Asp Thr Glu Asn Glu Glu Tyr 565570 575 His Ser Asp Glu Gln Asn Asp Thr Gln Lys Gln Phe Cys Glu Glu Gln580 585 590 Asn Thr Gly Ile Leu His Asp Glu Ile Leu Ile His Glu Glu LysGln 595 600 605 Ile Glu Val Val Glu Lys Met Asn Ser Glu Leu Ser Leu SerCys Lys 610 615 620 Lys Glu Lys Asp Ile Leu His Glu Asn Ser Thr Leu ArgGlu Glu Ile 625 630 635 640 Ala Met Leu Arg Leu Glu Leu Asp Thr Met LysHis Gln Ser Gln Leu 645 650 655 <210> SEQ ID NO 306 <211> LENGTH: 671<212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 306 Met ValVal Glu Val Asp Ser Met Pro Ala Ala Ser Ser Val Lys Lys 1 5 10 15 ProPhe Gly Leu Arg Ser Lys Met Gly Lys Trp Cys Cys Arg Cys Phe 20 25 30 ProCys Cys Arg Glu Ser Gly Lys Ser Asn Val Gly Thr Ser Gly Asp 35 40 45 HisAsp Asp Ser Ala Met Lys Thr Leu Arg Ser Lys Met Gly Lys Trp 50 55 60 CysArg His Cys Phe Pro Cys Cys Arg Gly Ser Gly Lys Ser Asn Val 65 70 75 80Gly Ala Ser Gly Asp His Asp Asp Ser Ala Met Lys Thr Leu Arg Asn 85 90 95Lys Met Gly Lys Trp Cys Cys His Cys Phe Pro Cys Cys Arg Gly Ser 100 105110 Gly Lys Ser Lys Val Gly Ala Trp Gly Asp Tyr Asp Asp Ser Ala Phe 115120 125 Met Glu Pro Arg Tyr His Val Arg Gly Glu Asp Leu Asp Lys Leu His130 135 140 Arg Ala Ala Trp Trp Gly Lys Val Pro Arg Lys Asp Leu Ile ValMet 145 150 155 160 Leu Arg Asp Thr Asp Val Asn Lys Lys Asp Lys Gln LysArg Thr Ala 165 170 175 Leu His Leu Ala Ser Ala Asn Gly Asn Ser Glu ValVal Lys Leu Leu 180 185 190 Leu Asp Arg Arg Cys Gln Leu Asn Val Leu AspAsn Lys Lys Arg Thr 195 200 205 Ala Leu Ile Lys Ala Val Gln Cys Gln GluAsp Glu Cys Ala Leu Met 210 215 220 Leu Leu Glu His Gly Thr Asp Pro AsnIle Pro Asp Glu Tyr Gly Asn 225 230 235 240 Thr Thr Leu His Tyr Ala IleTyr Asn Glu Asp Lys Leu Met Ala Lys 245 250 255 Ala Leu Leu Leu Tyr GlyAla Asp Ile Glu Ser Lys Asn Lys His Gly 260 265 270 Leu Thr Pro Leu LeuLeu Gly Val His Glu Gln Lys Gln Gln Val Val 275 280 285 Lys Phe Leu IleLys Lys Lys Ala Asn Leu Asn Ala Leu Asp Arg Tyr 290 295 300 Gly Arg ThrAla Leu Ile Leu Ala Val Cys Cys Gly Ser Ala Ser Ile 305 310 315 320 ValSer Leu Leu Leu Glu Gln Asn Ile Asp Val Ser Ser Gln Asp Leu 325 330 335Ser Gly Gln Thr Ala Arg Glu Tyr Ala Val Ser Ser His His His Val 340 345350 Ile Cys Gln Leu Leu Ser Asp Tyr Lys Glu Lys Gln Met Leu Lys Ile 355360 365 Ser Ser Glu Asn Ser Asn Pro Glu Gln Asp Leu Lys Leu Thr Ser Glu370 375 380 Glu Glu Ser Gln Arg Phe Lys Gly Ser Glu Asn Ser Gln Pro GluLys 385 390 395 400 Met Ser Gln Glu Pro Glu Ile Asn Lys Asp Gly Asp ArgGlu Val Glu 405 410 415 Glu Glu Met Lys Lys His Glu Ser Asn Asn Val GlyLeu Leu Glu Asn 420 425 430 Leu Thr Asn Gly Val Thr Ala Gly Asn Gly AspAsn Gly Leu Ile Pro 435 440 445 Gln Arg Lys Ser Arg Thr Pro Glu Asn GlnGln Phe Pro Asp Asn Glu 450 455 460 Ser Glu Glu Tyr His Arg Ile Cys GluLeu Val Ser Asp Tyr Lys Glu 465 470 475 480 Lys Gln Met Pro Lys Tyr SerSer Glu Asn Ser Asn Pro Glu Gln Asp 485 490 495 Leu Lys Leu Thr Ser GluGlu Glu Ser Gln Arg Leu Glu Gly Ser Glu 500 505 510 Asn Gly Gln Pro GluLys Arg Ser Gln Glu Pro Glu Ile Asn Lys Asp 515 520 525 Gly Asp Arg GluLeu Glu Asn Phe Met Ala Ile Glu Glu Met Lys Lys 530 535 540 His Gly SerThr His Val Gly Phe Pro Glu Asn Leu Thr Asn Gly Ala 545 550 555 560 ThrAla Gly Asn Gly Asp Asp Gly Leu Ile Pro Pro Arg Lys Ser Arg 565 570 575Thr Pro Glu Ser Gln Gln Phe Pro Asp Thr Glu Asn Glu Glu Tyr His 580 585590 Ser Asp Glu Gln Asn Asp Thr Gln Lys Gln Phe Cys Glu Glu Gln Asn 595600 605 Thr Gly Ile Leu His Asp Glu Ile Leu Ile His Glu Glu Lys Gln Ile610 615 620 Glu Val Val Glu Lys Met Asn Ser Glu Leu Ser Leu Ser Cys LysLys 625 630 635 640 Glu Lys Asp Ile Leu His Glu Asn Ser Thr Leu Arg GluGlu Ile Ala 645 650 655 Met Leu Arg Leu Glu Leu Asp Thr Met Lys His GlnSer Gln Leu 660 665 670 <210> SEQ ID NO 307 <211> LENGTH: 800 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 307 atkagcttccgcttctgaca acactagaga tccctcccct ccctcagggt atggccctcc 60 acttcatttttggtacataa catctttata ggacaggggt aaaatcccaa tactaacagg 120 agaatgcttaggactctaac aggtttttga gaatgtgttg gtaagggcca ctcaatccaa 180 tttttcttggtcctccttgt ggtctaggag gacaggcaag ggtgcagatt ttcaagaatg 240 catcagtaagggccactaaa tccgaccttc ctcgttcctc cttgtggtct gggaggaaaa 300 ctagtgtttctgttgctgtg tcagtgagca caactattcc gatcagcagg gtccagggac 360 cactgcaggttcttgggcag ggggagaaac aaaacaaacc aaaaccatgg gcrgttttgt 420 ctttcagatgggaaacactc aggcatcaac aggctcacct ttgaaatgca tcctaagcca 480 atgggacaaatttgacccac aaaccctgga aaaagaggtg gctcattttt tttgcactat 540 ggcttggccccaacattctc tctctgatgg ggaaaaatgg ccacctgagg gaagtacaga 600 ttacaatactatcctgcagc ttgacctttt ctgtaagagg gaaggcaaat ggagtgaaat 660 accttatgtccaagctttct tttcattgaa ggagaataca ctatgcaaag cttgaaattt 720 acatcccacaggaggacctc tcagcttacc cccatatcct agcctcccta tagctcccct 780 tcctattagtgataagcctc 800 <210> SEQ ID NO 308 <211> LENGTH: 102 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: VARIANT<222> LOCATION: 3 <223> OTHER INFORMATION: Xaa = Any Amino Acid <400>SEQUENCE: 308 Met Gly Xaa Phe Val Phe Gln Met Gly Asn Thr Gln Ala SerThr Gly 1 5 10 15 Ser Pro Leu Lys Cys Ile Leu Ser Gln Trp Asp Lys PheAsp Pro Gln 20 25 30 Thr Leu Glu Lys Glu Val Ala His Phe Phe Cys Thr MetAla Trp Pro 35 40 45 Gln His Ser Leu Ser Asp Gly Glu Lys Trp Pro Pro GluGly Ser Thr 50 55 60 Asp Tyr Asn Thr Ile Leu Gln Leu Asp Leu Phe Cys LysArg Glu Gly 65 70 75 80 Lys Trp Ser Glu Ile Pro Tyr Val Gln Ala Phe PheSer Leu Lys Glu 85 90 95 Asn Thr Leu Cys Lys Ala 100 <210> SEQ ID NO 309<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Made in the lab <400> SEQUENCE:309 Leu Met Ala Glu Glu Tyr Thr Ile Val 1 5 <210> SEQ ID NO 310 <211>LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Made in the lab <400> SEQUENCE: 310Lys Leu Met Ala Lys Ala Leu Leu Leu 1 5 <210> SEQ ID NO 311 <211>LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Made in the lab <400> SEQUENCE: 311Gly Leu Thr Pro Leu Leu Leu Gly Ile 1 5 <210> SEQ ID NO 312 <211>LENGTH: 10 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Made in the lab <400> SEQUENCE: 312Lys Leu Val Leu Asp Arg Arg Cys Gln Leu 1 5 10 <210> SEQ ID NO 313 <211>LENGTH: 1852 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 313 ggcacgagaa ttaaaaccct cagcaaaaca ggcatagaag ggacataccttaaagtaata 60 aaaaccacct atgacaagcc cacagccaac ataatactaa atggggaaaagttagaagca 120 tttcctctga gaactgcaac aataaataca aggatgctgg attttgtcaaatgccttttc 180 tgtgtctgtt gagatgctta tgtgactttg cttttaattc tgtttatgtgattatcacat 240 ttattgactt gcctgtgtta gaccggaaga gctggggtgt ttctcaggagccaccgtgtg 300 ctgcggcagc ttcgggataa cttgaggctg catcactggg gaagaaacacaytcctgtcc 360 gtggcgctga tggctgagga cagagcttca gtgtggcttc tctgcgactggcttcttcgg 420 ggagttcttc cttcatagtt catccatatg gctccagagg aaaattatattattttgtta 480 tggatgaaga gtattacgtt gtgcagatat actgcagtgt cttcatctcttgatgtgtga 540 ttgggtaggt tccaccatgt tgccgcagat gacatgattt cagtacctgtgtctggctga 600 aaagtgtttg tttgtgaatg gatattgtgg tttctggatc tcatcctctgtgggtggaca 660 gctttctcca ccttgctgga agtgacctgc tgtccagaag tttgatggctgaggagtata 720 ccatcgtgca tgcatctttc atttcctgca tttcttcctc cctggatggacagggggagc 780 ggcaagagca acgtgggcac ttctggagac cacaacgact cctctgtgaagacgcttggg 840 agcaagaggt gcaagtggtg ctgccactgc ttcccctgct gcagggggagcggcaagagc 900 aacgtggtcg cttggggaga ctacgatgac agcgccttca tggatcccaggtaccacgtc 960 catggagaag atctggacaa gctccacaga gctgcctggt ggggtaaagtccccagaaag 1020 gatctcatcg tcatgctcag ggacacggat gtgaacaaga gggacaagcaaaagaggact 1080 gctctacatc tggcctctgc caatgggaat tcagaagtag taaaactcgtgctggacaga 1140 cgatgtcaac ttaatgtcct tgacaacaaa aagaggacag ctctgacaaaggccgtacaa 1200 tgccaggaag atgaatgtgc gttaatgttg ctggaacatg gcactgatccaaatattcca 1260 gatgagtatg gaaataccac tctacactat gctgtctaca atgaagataaattaatggcc 1320 aaagcactgc tcttatacgg tgctgatatc gaatcaaaaa acaagcatggcctcacacca 1380 ctgctacttg gtatacatga gcaaaaacag caagtggtga aatttttaatcaagaaaaaa 1440 gcgaatttaa atgcgctgga tagatatgga agaactgctc tcatacttgctgtatgttgt 1500 ggatcagcaa gtatagtcag ccctctactt gagcaaaatg ttgatgtatcttctcaagat 1560 ctggaaagac ggccagagag tatgctgttt ctagtcatca tcatgtaatttgccagttac 1620 tttctgacta caaagaaaaa cagatgttaa aaatctcttc tgaaaacagcaatccagaac 1680 aagacttaaa gctgacatca gaggaagagt cacaaaggct taaaggaagtgaaaacagcc 1740 agccagagct agaagattta tggctattga agaagaatga agaacacggaagtactcatg 1800 tgggattccc agaaaacctg actaacggtg ccgctgctgg caatggtgatga 1852 <210> SEQ ID NO 314 <211> LENGTH: 879 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 314 atgcatcttt catttcctgcatttcttcct ccctggatgg acagggggag cggcaagagc 60 aacgtgggca cttctggagaccacaacgac tcctctgtga agacgcttgg gagcaagagg 120 tgcaagtggt gctgccactgcttcccctgc tgcaggggga gcggcaagag caacgtggtc 180 gcttggggag actacgatgacagcgccttc atggatccca ggtaccacgt ccatggagaa 240 gatctggaca agctccacagagctgcctgg tggggtaaag tccccagaaa ggatctcatc 300 gtcatgctca gggacacggatgtgaacaag agggacaagc aaaagaggac tgctctacat 360 ctggcctctg ccaatgggaattcagaagta gtaaaactcg tgctggacag acgatgtcaa 420 cttaatgtcc ttgacaacaaaaagaggaca gctctgacaa aggccgtaca atgccaggaa 480 gatgaatgtg cgttaatgttgctggaacat ggcactgatc caaatattcc agatgagtat 540 ggaaatacca ctctacactatgctgtctac aatgaagata aattaatggc caaagcactg 600 ctcttatacg gtgctgatatcgaatcaaaa aacaagcatg gcctcacacc actgctactt 660 ggtatacatg agcaaaaacagcaagtggtg aaatttttaa tcaagaaaaa agcgaattta 720 aatgcgctgg atagatatggaagaactgct ctcatacttg ctgtatgttg tggatcagca 780 agtatagtca gccctctacttgagcaaaat gttgatgtat cttctcaaga tctggaaaga 840 cggccagaga gtatgctgtttctagtcatc atcatgtaa 879 <210> SEQ ID NO 315 <211> LENGTH: 292 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 315 Met His LeuSer Phe Pro Ala Phe Leu Pro Pro Trp Met Asp Arg Gly 5 10 15 Ser Gly LysSer Asn Val Gly Thr Ser Gly Asp His Asn Asp Ser Ser 20 25 30 Val Lys ThrLeu Gly Ser Lys Arg Cys Lys Trp Cys Cys His Cys Phe 35 40 45 Pro Cys CysArg Gly Ser Gly Lys Ser Asn Val Val Ala Trp Gly Asp 50 55 60 Tyr Asp AspSer Ala Phe Met Asp Pro Arg Tyr His Val His Gly Glu 65 70 75 80 Asp LeuAsp Lys Leu His Arg Ala Ala Trp Trp Gly Lys Val Pro Arg 85 90 95 Lys AspLeu Ile Val Met Leu Arg Asp Thr Asp Val Asn Lys Arg Asp 100 105 110 LysGln Lys Arg Thr Ala Leu His Leu Ala Ser Ala Asn Gly Asn Ser 115 120 125Glu Val Val Lys Leu Val Leu Asp Arg Arg Cys Gln Leu Asn Val Leu 130 135140 Asp Asn Lys Lys Arg Thr Ala Leu Thr Lys Ala Val Gln Cys Gln Glu 145150 155 160 Asp Glu Cys Ala Leu Met Leu Leu Glu His Gly Thr Asp Pro AsnIle 165 170 175 Pro Asp Glu Tyr Gly Asn Thr Thr Leu His Tyr Ala Val TyrAsn Glu 180 185 190 Asp Lys Leu Met Ala Lys Ala Leu Leu Leu Tyr Gly AlaAsp Ile Glu 195 200 205 Ser Lys Asn Lys His Gly Leu Thr Pro Leu Leu LeuGly Ile His Glu 210 215 220 Gln Lys Gln Gln Val Val Lys Phe Leu Ile LysLys Lys Ala Asn Leu 225 230 235 240 Asn Ala Leu Asp Arg Tyr Gly Arg ThrAla Leu Ile Leu Ala Val Cys 245 250 255 Cys Gly Ser Ala Ser Ile Val SerPro Leu Leu Glu Gln Asn Val Asp 260 265 270 Val Ser Ser Gln Asp Leu GluArg Arg Pro Glu Ser Met Leu Phe Leu 275 280 285 Val Ile Ile Met 290<210> SEQ ID NO 316 <211> LENGTH: 584 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 316 agttgggcca aattcccctc cccctacagcttgaagggga cataaccaat agcctggggt 60 ttttttgtgg tcctttggag atttctttgcttattttctt ctgggtgggg gtgattagag 120 gaggcttatc actaatagga aggggagctatagggaggct aggatatggg ggtaagctga 180 gaggtcctcc tgtgggatgt aaatttcaagctttgcatag tgtattctcc ttcaatgaaa 240 agaaagcttg gacataaggt atttcactccatttgccttc cctcttacag aaaaggtcaa 300 gctgcaggat agtattgtaa tctgtacttccctcaggtgg ccatttttcc ccatcagaga 360 gagaatgttg gggccaagcc atagtgcagaaaaaaaaatg agccacctct ttttccaggg 420 tttgtgggtc aaatttgtcc cattggcttaggatgcattt caaaggtgag cctgttgatg 480 cctgagtgtt tcccatctga aagacaaaactgcccatggt tttggtttgt tttgtttctc 540 cccctgccca agaactatca aactcctgagccaacaacta aaaa 584 <210> SEQ ID NO 317 <211> LENGTH: 829 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 317 attagcttccgcttctgaca acactagaga tccctcccct ccctcagggt atggccctcc 60 acttcatttttggtacataa catctttata ggacaggggt aaaatcccaa tactaacagg 120 agaatgcttaggactctaac aggtttttga gaatgtgttg gtaagggcca ctcaatccaa 180 tttttcttggtcctccttgt ggtctaggag gacaggcaag ggtgcagatt ttcaagaatg 240 catcagtaagggccactaaa tccgaccttc ctcgttcctc cttgtggtct gggaggaaaa 300 ctagtgtttctgttgctgtg tcagtgagca caactattcc gatcagcagg gtccagggac 360 cactgcaggttcttgggcag ggggagaaac aaaacaaacc aaaaccatgg gcagttttgt 420 ctttcagatgggaaacactc aggcatcaac aggctcacct ttgaaatgca tcctaagcca 480 atgggacaaatttgacccac aaaccctgga aaaagaggtg gctcattttt tttgcactat 540 ggcttggccccaacattctc tctctgatgg ggaaaaatgg ccacctgagg gaagtacaga 600 ttacaatactatcctgcagc ttgacctttt ctgtaagagg gaaggcaaat ggagtgaaat 660 accttatgtccaagctttct tttcattgaa ggagaataca ctatgcaaag cttgaaattt 720 acatcccacaggaggacctc tcagcttacc cccatatcct agcctcccta tagctcccct 780 tcctattagtgataagcctc ctctaatcac ccccacccag aagaaaata 829 <210> SEQ ID NO 318 <211>LENGTH: 30 <212> TYPE: PRT <213> ORGANISM: Homo sapien <400> SEQUENCE:318 Thr Ala Ala Ser Asp Asn Phe Gln Leu Ser Gln Gly Gly Gln Gly Phe 1 510 15 Ala Ile Pro Ile Gly Gln Ala Met Ala Ile Ala Gly Gln Ile 20 25 30<210> SEQ ID NO 319 <211> LENGTH: 41 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer<400> SEQUENCE: 319 ggcctctgcc aatgggaact cagaagtagt aaaactcctg c 41<210> SEQ ID NO 320 <211> LENGTH: 41 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer<400> SEQUENCE: 320 gcaggagttt tactacttct gagttcccat tggcagaggc c 41<210> SEQ ID NO 321 <211> LENGTH: 60 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer<400> SEQUENCE: 321 ggggaattcc cgctggtgcc gcgcggcagc cctatggtggttgaggttga 50 tccatgccg 60 <210> SEQ ID NO 322 <211> LENGTH: 42 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: PCR primer <400> SEQUENCE: 322 cccgaattct tatttatttctggttcttga gacattttct gg 42 <210> SEQ ID NO 323 <211> LENGTH: 1590 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 323 atgcatcaccatcaccatca cacggccgcg tccgataact tccagctgtc ccagggtggg 60 cagggattcgccattccgat cgggcaggcg atggcgatcg cgggccagat caagcttccc 120 accgttcatatcgggcctac cgccttcctc ggcttgggtg ttgtcgacaa caacggcaac 180 ggcgcacgagtccaacgcgt ggtcgggagc gctccggcgg caagtctcgg catctccacc 240 ggcgacgtgatcaccgcggt cgacggcgct ccgatcaact cggccaccgc gatggcggac 300 gcgcttaacgggcatcatcc cggtgacgtc atctcggtga cctggcaaac caagtcgggc 360 ggcacgcgtacagggaacgt gacattggcc gagggacccc cggccgaatt cccgctggtg 420 ccgcgcggcagccctatggt ggttgaggtt gattccatgc cggctgcttc ttctgtgaag 480 aagccatttggtctcaggag caagatgggc aagtggtgct gccgttgctt cccctgctgc 540 agggagagcggcaagagcaa cgtgggcact tctggagacc acgacgactc tgctatgaag 600 acactcaggagcaagatggg caagtggtgc cgccactgct tcccctgctg cagggggagt 660 ggcaagagcaacgtgggcgc ttctggagac cacgacgact ctgctatgaa gacactcagg 720 aacaagatgggcaagtggtg ctgccactgc ttcccctgct gcagggggag cggcaagagc 780 aaggtgggcgcttggggaga ctacgatgac agygccttca tggagcccag gtaccacgtc 840 cgtggagaagatctggacaa gctccacaga gctgcctggt ggggtaaagt ccccagaaag 900 gatctcatcgtcatgctcag ggacactgac gtgaacaaga aggacaagca aaagaggact 960 gctctacatctggcctctgc caatgggaat tcagaagtag taaaactcct gctggacaga 1020 cgatgtcaacttaatgtcct tgacaacaaa aagaggacag ctctgataaa ggccgtacaa 1080 tgccaggaagatgaatgtgc gttaatgttg ctggaacatg gcactgatcc aaatattcca 1140 gatgagtatggaaataccac tctgcactac gctatctata atgaagataa attaatggcc 1200 aaagcactgctcttatatgg tgctgatatc gaatcaaaaa acaagcatgg cctcacacca 1260 ctgttacttggtgtacatga gcaaaaacag caagtcgtga aatttttaat caagaaaaaa 1320 gcgaatttaaatgcactgga tagatatgga aggactgctc tcatacttgc tgtatgttgt 1380 ggatcagcaagtatagtcag ccttctactt gagcaaaata ttgatgtatc ttctcaagat 1440 ctatctggacagacggccag agagtatgct gtttctagtc atcatcatgt aatttgccag 1500 ttactttctgactacaaaga aaaacagatg ctaaaaatct cttctgaaaa cagcaatcca 1560 gaaaatgtctcaagaaccag aaataaataa 1590 <210> SEQ ID NO 324 <211> LENGTH: 529 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 324 Met His HisHis His His His Thr Ala Ala Ser Asp Asn Phe Gln Leu 5 10 15 Ser Gln GlyGly Gln Gly Phe Ala Ile Pro Ile Gly Gln Ala Met Ala 20 25 30 Ile Ala GlyGln Ile Lys Leu Pro Thr Val His Ile Gly Pro Thr Ala 35 40 45 Phe Leu GlyLeu Gly Val Val Asp Asn Asn Gly Asn Gly Ala Arg Val 50 55 60 Gln Arg ValVal Gly Ser Ala Pro Ala Ala Ser Leu Gly Ile Ser Thr 65 70 75 80 Gly AspVal Ile Thr Ala Val Asp Gly Ala Pro Ile Asn Ser Ala Thr 85 90 95 Ala MetAla Asp Ala Leu Asn Gly His His Pro Gly Asp Val Ile Ser 100 105 110 ValThr Trp Gln Thr Lys Ser Gly Gly Thr Arg Thr Gly Asn Val Thr 115 120 125Leu Ala Glu Gly Pro Pro Ala Glu Phe Pro Leu Val Pro Arg Gly Ser 130 135140 Pro Met Val Val Glu Val Asp Ser Met Pro Ala Ala Ser Ser Val Lys 145150 155 160 Lys Pro Phe Gly Leu Arg Ser Lys Met Gly Lys Trp Cys Cys ArgCys 165 170 175 Phe Pro Cys Cys Arg Glu Ser Gly Lys Ser Asn Val Gly ThrSer Gly 180 185 190 Asp His Asp Asp Ser Ala Met Lys Thr Leu Arg Ser LysMet Gly Lys 195 200 205 Trp Cys Arg His Cys Phe Pro Cys Cys Arg Gly SerGly Lys Ser Asn 210 215 220 Val Gly Ala Ser Gly Asp His Asp Asp Ser AlaMet Lys Thr Leu Arg 225 230 235 240 Asn Lys Met Gly Lys Trp Cys Cys HisCys Phe Pro Cys Cys Arg Gly 245 250 255 Ser Gly Lys Ser Lys Val Gly AlaTrp Gly Asp Tyr Asp Asp Ser Ala 260 265 270 Phe Met Glu Pro Arg Tyr HisVal Arg Gly Glu Asp Leu Asp Lys Leu 275 280 285 His Arg Ala Ala Trp TrpGly Lys Val Pro Arg Lys Asp Leu Ile Val 290 295 300 Met Leu Arg Asp ThrAsp Val Asn Lys Lys Asp Lys Gln Lys Arg Thr 305 310 315 320 Ala Leu HisLeu Ala Ser Ala Asn Gly Asn Ser Glu Val Val Lys Leu 325 330 335 Leu LeuAsp Arg Arg Cys Gln Leu Asn Val Leu Asp Asn Lys Lys Arg 340 345 350 ThrAla Leu Ile Lys Ala Val Gln Cys Gln Glu Asp Glu Cys Ala Leu 355 360 365Met Leu Leu Glu His Gly Thr Asp Pro Asn Ile Pro Asp Glu Tyr Gly 370 375380 Asn Thr Thr Leu His Tyr Ala Ile Tyr Asn Glu Asp Lys Leu Met Ala 385390 395 400 Lys Ala Leu Leu Leu Tyr Gly Ala Asp Ile Glu Ser Lys Asn LysHis 405 410 415 Gly Leu Thr Pro Leu Leu Leu Gly Val His Glu Gln Lys GlnGln Val 420 425 430 Val Lys Phe Leu Ile Lys Lys Lys Ala Asn Leu Asn AlaLeu Asp Arg 435 440 445 Tyr Gly Arg Thr Ala Leu Ile Leu Ala Val Cys CysGly Ser Ala Ser 450 455 460 Ile Val Ser Leu Leu Leu Glu Gln Asn Ile AspVal Ser Ser Gln Asp 465 470 475 480 Leu Ser Gly Gln Thr Ala Arg Glu TyrAla Val Ser Ser His His His 485 490 495 Val Ile Cys Gln Leu Leu Ser AspTyr Lys Glu Lys Gln Met Leu Lys 500 505 510 Ile Ser Ser Glu Asn Ser AsnPro Glu Asn Val Ser Arg Thr Arg Asn 515 520 525 Lys <210> SEQ ID NO 325<211> LENGTH: 1155 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 325 atggtggctg aggtttgttc aatgcccact gcctctactg tgaagaagccatttgatctc 60 aggagcaaga tgggcaagtg gtgccaccac cgcttcccct gctgcagggggagcggcaag 120 agcaacatgg gcacttctgg agaccacgac gactccttta tgaagatgctcaggagcaag 180 atgggcaagt gttgccgcca ctgcttcccc tgctgcaggg ggagcggcacgagcaacgtg 240 ggcacttctg gagaccatga aaactccttt atgaagatgc tcaggagcaagatgggcaag 300 tggtgctgtc actgcttccc ctgctgcagg gggagcggca agagcaacgtgggcgcttgg 360 ggagactacg accacagcgc cttcatggag ccgaggtacc acatccgtcgagaagatctg 420 gacaagctcc acagagctgc ctggtggggt aaagtcccca gaaaggatctcatcgtcatg 480 ctcagggaca ctgacatgaa caagagggac aaggaaaaga ggactgctctacatttggcc 540 tctgccaatg gaaattcaga agtagtacaa ctcctgctgg acagacgatgtcaacttaat 600 gtccttgaca acaaaaaaag gacagctctg ataaaggcca tacaatgccaggaagatgaa 660 tgtgtgttaa tgttgctgga acatggcgct gatcgaaata ttccagatgagtatggaaat 720 accgctctac actatgctat ctacaatgaa gataaattaa tggccaaagcactgctctta 780 tatggtgctg atattgaatc aaaaaacaag gttggcctca caccacttttgcttggcgta 840 catgaacaaa aacagcaagt ggtgaaattt ttaatcaaga aaaaagctaatttaaatgta 900 cttgatagat atggaaggac tgccctcata cttgctgtat gttgtggatcagcaagtata 960 gtcaatcttc tacttgagca aaatgttgat gtatcttctc aagatctatctggacagacg 1020 gccagagagt atgctgtttc tagtcatcat catgtaattt gtgaattactttctgactat 1080 aaagaaaaac agatgctaaa aatctcttct gaaaacagca atccagaaaatgtctcaaga 1140 accagaaata aataa 1155 <210> SEQ ID NO 326 <211> LENGTH:384 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 326 MetVal Ala Glu Val Cys Ser Met Pro Thr Ala Ser Thr Val Lys Lys 5 10 15 ProPhe Asp Leu Arg Ser Lys Met Gly Lys Trp Cys His His Arg Phe 20 25 30 ProCys Cys Arg Gly Ser Gly Lys Ser Asn Met Gly Thr Ser Gly Asp 35 40 45 HisAsp Asp Ser Phe Met Lys Met Leu Arg Ser Lys Met Gly Lys Cys 50 55 60 CysArg His Cys Phe Pro Cys Cys Arg Gly Ser Gly Thr Ser Asn Val 65 70 75 80Gly Thr Ser Gly Asp His Glu Asn Ser Phe Met Lys Met Leu Arg Ser 85 90 95Lys Met Gly Lys Trp Cys Cys His Cys Phe Pro Cys Cys Arg Gly Ser 100 105110 Gly Lys Ser Asn Val Gly Ala Trp Gly Asp Tyr Asp His Ser Ala Phe 115120 125 Met Glu Pro Arg Tyr His Ile Arg Arg Glu Asp Leu Asp Lys Leu His130 135 140 Arg Ala Ala Trp Trp Gly Lys Val Pro Arg Lys Asp Leu Ile ValMet 145 150 155 160 Leu Arg Asp Thr Asp Met Asn Lys Arg Asp Lys Glu LysArg Thr Ala 165 170 175 Leu His Leu Ala Ser Ala Asn Gly Asn Ser Glu ValVal Gln Leu Leu 180 185 190 Leu Asp Arg Arg Cys Gln Leu Asn Val Leu AspAsn Lys Lys Arg Thr 195 200 205 Ala Leu Ile Lys Ala Ile Gln Cys Gln GluAsp Glu Cys Val Leu Met 210 215 220 Leu Leu Glu His Gly Ala Asp Arg AsnIle Pro Asp Glu Tyr Gly Asn 225 230 235 240 Thr Ala Leu His Tyr Ala IleTyr Asn Glu Asp Lys Leu Met Ala Lys 245 250 255 Ala Leu Leu Leu Tyr GlyAla Asp Ile Glu Ser Lys Asn Lys Val Gly 260 265 270 Leu Thr Pro Leu LeuLeu Gly Val His Glu Gln Lys Gln Gln Val Val 275 280 285 Lys Phe Leu IleLys Lys Lys Ala Asn Leu Asn Val Leu Asp Arg Tyr 290 295 300 Gly Arg ThrAla Leu Ile Leu Ala Val Cys Cys Gly Ser Ala Ser Ile 305 310 315 320 ValAsn Leu Leu Leu Glu Gln Asn Val Asp Val Ser Ser Gln Asp Leu 325 330 335Ser Gly Gln Thr Ala Arg Glu Tyr Ala Val Ser Ser His His His Val 340 345350 Ile Cys Glu Leu Leu Ser Asp Tyr Lys Glu Lys Gln Met Leu Lys Ile 355360 365 Ser Ser Glu Asn Ser Asn Pro Glu Asn Val Ser Arg Thr Arg Asn Lys370 375 380 <210> SEQ ID NO 327 <211> LENGTH: 634 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 327 gactgctcta catctggcctctgccaatgg aaattcagaa gtagtaaaac tcctgctgga 60 cagacgatgt caacttaatatccttgacaa caaaaagagg acagctctga caaaggccgt 120 acaatgccag gaagatgaatgtgcgttaat gttgctggaa catggcactg atccgaatat 180 tccagatgag tatggaaataccgctctaca ctatgctatc tacaatgaag ataaattaat 240 ggccaaagca ctgctcttatacggtgctga tatcgaatca aaaaacaagc atggcctcac 300 accactgtta cttggtgtacatgagcaaaa acagcaagtg gtgaaatttt taatcaagaa 360 aaaagcaaat ttaaatgcactggatagata tggaagaact gctctcatac ttgctgtatg 420 ttgtggatcg gcaagtatagtcagccttct acttgagcaa aacattgatg tatcttctca 480 agatctatct ggacagacggccagagagta tgctgtttct agtcgtcata atgtaatttg 540 ccagttactt tctgactacaaagaaaaaca gatactaaaa gtctcttctg aaaacagcaa 600 tccaggaaat gtctcaagaaccagaaataa ataa 634 <210> SEQ ID NO 328 <211> LENGTH: 1155 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 328 atggtggttgaggttgattc catgccggct gcctcttctg tgaagaagcc atttggtctc 60 aggagcaagatgggcaagtg gtgctgccgt tgcttcccct gctgcaggga gagcggcaag 120 agcaacgtgggcacttctgg agaccacgac gactctgcta tgaagacact caggagcaag 180 atgggcaagtggtgccgcca ctgcttcccc tgctgcaggg ggagtggcaa gagcaacgtg 240 ggcgcttctggagaccacga cgactctgct atgaagacac tcaggaacaa gatgggcaag 300 tggtgctgccactgcttccc ctgctgcagg gggagcagca agagcaaggt gggcgcttgg 360 ggagactacgatgacagtgc cttcatggag cccaggtacc acgtccgtgg agaagatctg 420 gacaagctccacagagctgc ctggtggggt aaagtcccca gaaaggatct catcgtcatg 480 ctcagggacactgacgtgaa caagcaggac aagcaaaaga ggactgctct acatctggcc 540 tctgccaatgggaattcaga agtagtaaaa ctcctgctgg acagacgatg tcaacttaat 600 gtccttgacaacaaaaagag gacagctctg ataaaggccg tacaatgcca ggaagatgaa 660 tgtgcgttaatgttgctgga acatggcact gatccaaata ttccagatga gtatggaaat 720 accactctgcactacgctat ctataatgaa gataaattaa tggccaaagc actgctctta 780 tatggtgctgatatcgaatc aaaaaacaag catggcctca caccactgtt acttggtgta 840 catgagcaaaaacagcaagt cgtgaaattt ttaattaaga aaaaagcgaa tttaaatgca 900 ctggatagatatggaaggac tgctctcata cttgctgtat gttgtggatc agcaagtata 960 gtcagccttctacttgagca aaatattgat gtatcttctc aagatctatc tggacagacg 1020 gccagagagtatgctgtttc tagtcatcat catgtaattt gccagttact ttctgactac 1080 aaagaaaaacagatgctaaa aatctcttct gaaaacagca atccagaaaa tgtctcaaga 1140 accagaaataaataa 1155 <210> SEQ ID NO 329 <211> LENGTH: 1155 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 329 atggtggctg aggtttgttcaatgcccgct gcctctgctg tgaagaagcc atttgatctc 60 aggagcaaga tgggcaagtggtgccaccac cgcttcccct gctgcagggg gagcggcaag 120 agcaacatgg gcacttctggagaccacgac gactccttta tgaagacgct caggagcaag 180 atgggcaagt gttgccaccactgcttcccc tgctgcaggg ggagcggcac gagcaatgtg 240 ggcacttctg gagaccatgacaactccttt atgaagacac tcaggagcaa gatgggcaag 300 tggtgctgtc actgcttcccctgctgcagg gggagcggca agagcaacgt gggcacttgg 360 ggagactacg acgacagcgccttcatggag ccgaggtacc acgtccgtcg agaagatctg 420 gacaagctcc acagagctgcctggtggggt aaagtcccca gaaaggatct catcgtcatg 480 ctcagggaca ctgacatgaacaagagggac aagcaaaaga ggactgctct acatttggcc 540 tctgccaatg gaaattcagaagtagtacaa ctcctgctgg acagacgatg tcaacttaac 600 gtccttgaca acaaaaaaaggacagctctg ataaaggccg tacaatgcca ggaagatgaa 660 tgtgtgttaa tgttgctggaacatggcgct gatggaaata ttcaagatga gtatggaaat 720 accgctctac actatgctatctacaatgaa gataaattaa tggccaaagc actgctctta 780 tatggtgctg atattgaatcaaaaaacaag tgtggcctca caccactttt gcttggcgta 840 catgaacaaa aacagcaagtggtgaaattt ttaatcaaga aaaaagctaa tttaaatgca 900 cttgatagat atggaagaactgccctcata cttgctgtat gttgtggatc agcaagtata 960 gtcaatcttc tacttgagcaaaatgttgat gtatcttctc aagatctatc tggacagacg 1020 gccagagagt atgctgtttctagtcatcat catgtaattt gtgaattact ttctgactat 1080 aaagaaaaac agatgctaaaaatctcttct gaaaacagca atccagaaaa tgtctcaaga 1140 accagaaata aataa 1155<210> SEQ ID NO 330 <211> LENGTH: 1155 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 330 atggtggctg aggtttgttc aatgcccactgcctctactg tgaagaagcc atttgatctc 60 aggagcaaga tgggcaagtg gtgccaccaccgcttcccct gctgcagggg gagcggcaag 120 agcaacatgg gcacttctgg agaccacgacgactccttta tgaagatgct caggagcaag 180 atgggcaagt gttgccgcca ctgcttcccctgctgcaggg ggagcggcac gagcaacgtg 240 ggcacttctg gagaccatga aaactcctttatgaagatgc tcaggagcaa gatgggcaag 300 tggtgctgtc actgcttccc ctgctgcagggggagcggca agagcaacgt gggcgcttgg 360 ggagactacg accacagcgc cttcatggagccgaggtacc acatccgtcg agaagatctg 420 gacaagctcc acagagctgc ctggtggggtaaagtcccca gaaaggatct catcgtcatg 480 ctcagggaca ctgacatgaa caagagggacaaggaaaaga ggactgctct acatttggcc 540 tctgccaatg gaaattcaga agtagtacaactcctgctgg acagacgatg tcaacttaat 600 gtccttgaca acaaaaaaag gacagctctgataaaggcca tacaatgcca ggaagatgaa 660 tgtgtgttaa tgttgctgga acatggcgctgatcgaaata ttccagatga gtatggaaat 720 accgctctac actatgctat ctacaatgaagataaattaa tggccaaagc actgctctta 780 tatggtgctg atattgaatc aaaaaacaagtgtggcctca caccactttt gcttggcgta 840 catgaacaaa aacagcaagt ggtgaaatttttaatcaaga aaaaagctaa tttaaatgta 900 cttgatagat atggaagaac tgccctcatacttgctgtat gttgtggatc agcaagtata 960 gtcaatcttc tacttgagca aaatgttgatgtatcttctc aagatctatc tggacagacg 1020 gccagagagt atgctgtttc tagtcatcatcatgtaattt gtgaattact ttctgactat 1080 aaagaaaaac agatgctaaa aatctcttctgaaaacagca atccagaaaa tgtctcaaga 1140 accagaaata aataa 1155 <210> SEQ IDNO 331 <211> LENGTH: 210 <212> TYPE: PRT <213> ORGANISM: Homo sapiens<400> SEQUENCE: 331 Thr Ala Leu His Leu Ala Ser Ala Asn Gly Asn Ser GluVal Val Lys 5 10 15 Leu Leu Leu Asp Arg Arg Cys Gln Leu Asn Ile Leu AspAsn Lys Lys 20 25 30 Arg Thr Ala Leu Thr Lys Ala Val Gln Cys Gln Glu AspGlu Cys Ala 35 40 45 Leu Met Leu Leu Glu His Gly Thr Asp Pro Asn Ile ProAsp Glu Tyr 50 55 60 Gly Asn Thr Ala Leu His Tyr Ala Ile Tyr Asn Glu AspLys Leu Met 65 70 75 80 Ala Lys Ala Leu Leu Leu Tyr Gly Ala Asp Ile GluSer Lys Asn Lys 85 90 95 His Gly Leu Thr Pro Leu Leu Leu Gly Val His GluGln Lys Gln Gln 100 105 110 Val Val Lys Phe Leu Ile Lys Lys Lys Ala AsnLeu Asn Ala Leu Asp 115 120 125 Arg Tyr Gly Arg Thr Ala Leu Ile Leu AlaVal Cys Cys Gly Ser Ala 130 135 140 Ser Ile Val Ser Leu Leu Leu Glu GlnAsn Ile Asp Val Ser Ser Gln 145 150 155 160 Asp Leu Ser Gly Gln Thr AlaArg Glu Tyr Ala Val Ser Ser Arg His 165 170 175 Asn Val Ile Cys Gln LeuLeu Ser Asp Tyr Lys Glu Lys Gln Ile Leu 180 185 190 Lys Val Ser Ser GluAsn Ser Asn Pro Gly Asn Val Ser Arg Thr Arg 195 200 205 Asn Lys 210<210> SEQ ID NO 332 <211> LENGTH: 384 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 332 Met Val Ala Glu Val Cys Ser Met Pro ThrAla Ser Thr Val Lys Lys 5 10 15 Pro Phe Asp Leu Arg Ser Lys Met Gly LysTrp Cys His His Arg Phe 20 25 30 Pro Cys Cys Arg Gly Ser Gly Lys Ser AsnMet Gly Thr Ser Gly Asp 35 40 45 His Asp Asp Ser Phe Met Lys Met Leu ArgSer Lys Met Gly Lys Cys 50 55 60 Cys Arg His Cys Phe Pro Cys Cys Arg GlySer Gly Thr Ser Asn Val 65 70 75 80 Gly Thr Ser Gly Asp His Glu Asn SerPhe Met Lys Met Leu Arg Ser 85 90 95 Lys Met Gly Lys Trp Cys Cys His CysPhe Pro Cys Cys Arg Gly Ser 100 105 110 Gly Lys Ser Asn Val Gly Ala TrpGly Asp Tyr Asp His Ser Ala Phe 115 120 125 Met Glu Pro Arg Tyr His IleArg Arg Glu Asp Leu Asp Lys Leu His 130 135 140 Arg Ala Ala Trp Trp GlyLys Val Pro Arg Lys Asp Leu Ile Val Met 145 150 155 160 Leu Arg Asp ThrAsp Met Asn Lys Arg Asp Lys Glu Lys Arg Thr Ala 165 170 175 Leu His LeuAla Ser Ala Asn Gly Asn Ser Glu Val Val Gln Leu Leu 180 185 190 Leu AspArg Arg Cys Gln Leu Asn Val Leu Asp Asn Lys Lys Arg Thr 195 200 205 AlaLeu Ile Lys Ala Ile Gln Cys Gln Glu Asp Glu Cys Val Leu Met 210 215 220Leu Leu Glu His Gly Ala Asp Arg Asn Ile Pro Asp Glu Tyr Gly Asn 225 230235 240 Thr Ala Leu His Tyr Ala Ile Tyr Asn Glu Asp Lys Leu Met Ala Lys245 250 255 Ala Leu Leu Leu Tyr Gly Ala Asp Ile Glu Ser Lys Asn Lys CysGly 260 265 270 Leu Thr Pro Leu Leu Leu Gly Val His Glu Gln Lys Gln GlnVal Val 275 280 285 Lys Phe Leu Ile Lys Lys Lys Ala Asn Leu Asn Val LeuAsp Arg Tyr 290 295 300 Gly Arg Thr Ala Leu Ile Leu Ala Val Cys Cys GlySer Ala Ser Ile 305 310 315 320 Val Asn Leu Leu Leu Glu Gln Asn Val AspVal Ser Ser Gln Asp Leu 325 330 335 Ser Gly Gln Thr Ala Arg Glu Tyr AlaVal Ser Ser His His His Val 340 345 350 Ile Cys Glu Leu Leu Ser Asp TyrLys Glu Lys Gln Met Leu Lys Ile 355 360 365 Ser Ser Glu Asn Ser Asn ProGlu Asn Val Ser Arg Thr Arg Asn Lys 370 375 380 <210> SEQ ID NO 333<211> LENGTH: 384 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 333 Met Val Ala Glu Val Cys Ser Met Pro Ala Ala Ser Ala ValLys Lys 5 10 15 Pro Phe Asp Leu Arg Ser Lys Met Gly Lys Trp Cys His HisArg Phe 20 25 30 Pro Cys Cys Arg Gly Ser Gly Lys Ser Asn Met Gly Thr SerGly Asp 35 40 45 His Asp Asp Ser Phe Met Lys Thr Leu Arg Ser Lys Met GlyLys Cys 50 55 60 Cys His His Cys Phe Pro Cys Cys Arg Gly Ser Gly Thr SerAsn Val 65 70 75 80 Gly Thr Ser Gly Asp His Asp Asn Ser Phe Met Lys ThrLeu Arg Ser 85 90 95 Lys Met Gly Lys Trp Cys Cys His Cys Phe Pro Cys CysArg Gly Ser 100 105 110 Gly Lys Ser Asn Val Gly Thr Trp Gly Asp Tyr AspAsp Ser Ala Phe 115 120 125 Met Glu Pro Arg Tyr His Val Arg Arg Glu AspLeu Asp Lys Leu His 130 135 140 Arg Ala Ala Trp Trp Gly Lys Val Pro ArgLys Asp Leu Ile Val Met 145 150 155 160 Leu Arg Asp Thr Asp Met Asn LysArg Asp Lys Gln Lys Arg Thr Ala 165 170 175 Leu His Leu Ala Ser Ala AsnGly Asn Ser Glu Val Val Gln Leu Leu 180 185 190 Leu Asp Arg Arg Cys GlnLeu Asn Val Leu Asp Asn Lys Lys Arg Thr 195 200 205 Ala Leu Ile Lys AlaVal Gln Cys Gln Glu Asp Glu Cys Val Leu Met 210 215 220 Leu Leu Glu HisGly Ala Asp Gly Asn Ile Gln Asp Glu Tyr Gly Asn 225 230 235 240 Thr AlaLeu His Tyr Ala Ile Tyr Asn Glu Asp Lys Leu Met Ala Lys 245 250 255 AlaLeu Leu Leu Tyr Gly Ala Asp Ile Glu Ser Lys Asn Lys Cys Gly 260 265 270Leu Thr Pro Leu Leu Leu Gly Val His Glu Gln Lys Gln Gln Val Val 275 280285 Lys Phe Leu Ile Lys Lys Lys Ala Asn Leu Asn Ala Leu Asp Arg Tyr 290295 300 Gly Arg Thr Ala Leu Ile Leu Ala Val Cys Cys Gly Ser Ala Ser Ile305 310 315 320 Val Asn Leu Leu Leu Glu Gln Asn Val Asp Val Ser Ser GlnAsp Leu 325 330 335 Ser Gly Gln Thr Ala Arg Glu Tyr Ala Val Ser Ser HisHis His Val 340 345 350 Ile Cys Glu Leu Leu Ser Asp Tyr Lys Glu Lys GlnMet Leu Lys Ile 355 360 365 Ser Ser Glu Asn Ser Asn Pro Glu Asn Val SerArg Thr Arg Asn Lys 370 375 380 <210> SEQ ID NO 334 <211> LENGTH: 384<212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 334 Met ValVal Glu Val Asp Ser Met Pro Ala Ala Ser Ser Val Lys Lys 5 10 15 Pro PheGly Leu Arg Ser Lys Met Gly Lys Trp Cys Cys Arg Cys Phe 20 25 30 Pro CysCys Arg Glu Ser Gly Lys Ser Asn Val Gly Thr Ser Gly Asp 35 40 45 His AspAsp Ser Ala Met Lys Thr Leu Arg Ser Lys Met Gly Lys Trp 50 55 60 Cys ArgHis Cys Phe Pro Cys Cys Arg Gly Ser Gly Lys Ser Asn Val 65 70 75 80 GlyAla Ser Gly Asp His Asp Asp Ser Ala Met Lys Thr Leu Arg Asn 85 90 95 LysMet Gly Lys Trp Cys Cys His Cys Phe Pro Cys Cys Arg Gly Ser 100 105 110Ser Lys Ser Lys Val Gly Ala Trp Gly Asp Tyr Asp Asp Ser Ala Phe 115 120125 Met Glu Pro Arg Tyr His Val Arg Gly Glu Asp Leu Asp Lys Leu His 130135 140 Arg Ala Ala Trp Trp Gly Lys Val Pro Arg Lys Asp Leu Ile Val Met145 150 155 160 Leu Arg Asp Thr Asp Val Asn Lys Gln Asp Lys Gln Lys ArgThr Ala 165 170 175 Leu His Leu Ala Ser Ala Asn Gly Asn Ser Glu Val ValLys Leu Leu 180 185 190 Leu Asp Arg Arg Cys Gln Leu Asn Val Leu Asp AsnLys Lys Arg Thr 195 200 205 Ala Leu Ile Lys Ala Val Gln Cys Gln Glu AspGlu Cys Ala Leu Met 210 215 220 Leu Leu Glu His Gly Thr Asp Pro Asn IlePro Asp Glu Tyr Gly Asn 225 230 235 240 Thr Thr Leu His Tyr Ala Ile TyrAsn Glu Asp Lys Leu Met Ala Lys 245 250 255 Ala Leu Leu Leu Tyr Gly AlaAsp Ile Glu Ser Lys Asn Lys His Gly 260 265 270 Leu Thr Pro Leu Leu LeuGly Val His Glu Gln Lys Gln Gln Val Val 275 280 285 Lys Phe Leu Ile LysLys Lys Ala Asn Leu Asn Ala Leu Asp Arg Tyr 290 295 300 Gly Arg Thr AlaLeu Ile Leu Ala Val Cys Cys Gly Ser Ala Ser Ile 305 310 315 320 Val SerLeu Leu Leu Glu Gln Asn Ile Asp Val Ser Ser Gln Asp Leu 325 330 335 SerGly Gln Thr Ala Arg Glu Tyr Ala Val Ser Ser His His His Val 340 345 350Ile Cys Gln Leu Leu Ser Asp Tyr Lys Glu Lys Gln Met Leu Lys Ile 355 360365 Ser Ser Glu Asn Ser Asn Pro Glu Asn Val Ser Arg Thr Arg Asn Lys 370375 380 <210> SEQ ID NO 335 <211> LENGTH: 1185 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 335 atggtggttg aggttgattccatgccggct gcctcttctg tgaagaagcc atttggtctc 60 aggagcaaga tgggcaagtggtgctgccgt tgcttcccct gctgcaggga gagcggcaag 120 agcaacgtgg gcacttctggagaccacgac gactctgcta tgaagacact caggagcaag 180 atgggcaagt ggtgccgccactgcttcccc tgctgcaggg ggagtggcaa gagcaacgtg 240 ggcgcttctg gagaccacgacgactctgct atgaagacac tcaggaacaa gatgggcaag 300 tggtgctgcc actgcttcccctgctgcagg gggagcggca agagcaaggt gggcgcttgg 360 ggagactacg atgacagtgccttcatggag cccaggtacc acgtccgtgg agaagatctg 420 gacaagctcc acagagctgcctggtggggt aaagtcccca gaaaggatct catcgtcatg 480 ctcagggaca ctgacgtgaacaagaaggac aagcaaaaga ggactgctct acatctggcc 540 tctgccaatg ggaattcagaagtagtaaaa ctcctgctgg acagacgatg tcaacttaat 600 gtccttgaca acaaaaagaggacagctctg ataaaggccg tacaatgcca ggaagatgaa 660 tgtgcgttaa tgttgctggaacatggcact gatccaaata ttccagatga gtatggaaat 720 accactctgc actacgctatctataatgaa gataaattaa tggccaaagc actgctctta 780 tatggtgctg atatcgaatcaaaaaacaag catggcctca caccactgtt acttggtgta 840 catgagcaaa aacagcaagtcgtgaaattt ttaatcaaga aaaaagcgaa tttaaatgca 900 ctggatagat atggaaggactgctctcata cttgctgtat gttgtggatc agcaagtata 960 gtcagccttc tacttgagcaaaatattgat gtatcttctc aagatctatc tggacagacg 1020 gccagagagt atgctgtttctagtcatcat catgtaattt gccagttact ttctgactac 1080 aaagaaaaac agatgctaaaaatctcttct gaaaacagca atccagaaaa tgtctcaaga 1140 accagaaata aacatcatcaccatcatcat caccatcacc attaa 1185 <210> SEQ ID NO 336 <211> LENGTH: 394<212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 336 Met ValVal Glu Val Asp Ser Met Pro Ala Ala Ser Ser Val Lys Lys 5 10 15 Pro PheGly Leu Arg Ser Lys Met Gly Lys Trp Cys Cys Arg Cys Phe 20 25 30 Pro CysCys Arg Glu Ser Gly Lys Ser Asn Val Gly Thr Ser Gly Asp 35 40 45 His AspAsp Ser Ala Met Lys Thr Leu Arg Ser Lys Met Gly Lys Trp 50 55 60 Cys ArgHis Cys Phe Pro Cys Cys Arg Gly Ser Gly Lys Ser Asn Val 65 70 75 80 GlyAla Ser Gly Asp His Asp Asp Ser Ala Met Lys Thr Leu Arg Asn 85 90 95 LysMet Gly Lys Trp Cys Cys His Cys Phe Pro Cys Cys Arg Gly Ser 100 105 110Gly Lys Ser Lys Val Gly Ala Trp Gly Asp Tyr Asp Asp Ser Ala Phe 115 120125 Met Glu Pro Arg Tyr His Val Arg Gly Glu Asp Leu Asp Lys Leu His 130135 140 Arg Ala Ala Trp Trp Gly Lys Val Pro Arg Lys Asp Leu Ile Val Met145 150 155 160 Leu Arg Asp Thr Asp Val Asn Lys Lys Asp Lys Gln Lys ArgThr Ala 165 170 175 Leu His Leu Ala Ser Ala Asn Gly Asn Ser Glu Val ValLys Leu Leu 180 185 190 Leu Asp Arg Arg Cys Gln Leu Asn Val Leu Asp AsnLys Lys Arg Thr 195 200 205 Ala Leu Ile Lys Ala Val Gln Cys Gln Glu AspGlu Cys Ala Leu Met 210 215 220 Leu Leu Glu His Gly Thr Asp Pro Asn IlePro Asp Glu Tyr Gly Asn 225 230 235 240 Thr Thr Leu His Tyr Ala Ile TyrAsn Glu Asp Lys Leu Met Ala Lys 245 250 255 Ala Leu Leu Leu Tyr Gly AlaAsp Ile Glu Ser Lys Asn Lys His Gly 260 265 270 Leu Thr Pro Leu Leu LeuGly Val His Glu Gln Lys Gln Gln Val Val 275 280 285 Lys Phe Leu Ile LysLys Lys Ala Asn Leu Asn Ala Leu Asp Arg Tyr 290 295 300 Gly Arg Thr AlaLeu Ile Leu Ala Val Cys Cys Gly Ser Ala Ser Ile 305 310 315 320 Val SerLeu Leu Leu Glu Gln Asn Ile Asp Val Ser Ser Gln Asp Leu 325 330 335 SerGly Gln Thr Ala Arg Glu Tyr Ala Val Ser Ser His His His Val 340 345 350Ile Cys Gln Leu Leu Ser Asp Tyr Lys Glu Lys Gln Met Leu Lys Ile 355 360365 Ser Ser Glu Asn Ser Asn Pro Glu Asn Val Ser Arg Thr Arg Asn Lys 370375 380 His His His His His His His His His His 385 390 <210> SEQ ID NO337 <211> LENGTH: 34 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 337cggcggatcc accatggtgg ttgaggttga ttcc 34 <210> SEQ ID NO 338 <211>LENGTH: 74 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 338cggctctaga ttaatggtga tggtgatgat gatggtgatg atgtttattt ctggttcttg 60agacattttc tgga 74 <210> SEQ ID NO 339 <211> LENGTH: 1166 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 339 atggtggctgaggctggttc aatgccggct gcctcctctg tgaagaagcc atttggtctc 60 agaagcaagatgggcaagtg gtgccgccac tgcttcccct ggtgcagggg gagcggcaag 120 agcaacgtgggcacttctgg agaccacgac gattctgcta tgaagacact caggagcaag 180 atgggcaagtggtgccgcca ctgcttcccc tggtgcaggg ggagcagcaa gagcaacgtg 240 ggcacttctggagaccacga cgactctgct atgaagacac tcaggagcaa gatgggcaag 300 tggtgctgccactgcttccc ctgctgcagg gggagcggca agagcaaagt gggcccttgg 360 ggagactacgacgacagcgc tttcatggag ccgaggtacc acgtccgtcg agaagatctg 420 gacaagctccacagagctgc ctggtggggt aaagtcccca gaaaggatct catcgtcatg 480 ctcaaggacactgacatgaa caagaaggac aagcaaaaga ggactgctct acatctggcc 540 tctgccaatggaaattcaga agtagtaaaa ctcctgctgg acagacgatg tcaacttaat 600 atccttgacaacaaaaagag gacagctctg acaaaggccg tacaatgccg ggaagatgaa 660 tgtgcgttaatgttgctgga acatggcact gatccgaata ttccagatga gtatggaaat 720 accgctctacactatgctat ctacaatgaa gataaattaa tggccaaagc actgctctta 780 tacggtgctgatatcgaatc aaaaaacaag catggcctca caccactgtt acttggtgta 840 catgagcaaaaacagcaagt ggtgaaattc ttaatcaaga aaaaagcaaa tttaaatgca 900 ctggatagatatggaagaac tgctctcata cttgctgtat gttgtggatc ggcaagtata 960 gtcagccttctacttgagca aaacattgat gtatcttctc aagatctatc tggacagacg 1020 gccagagagtatgctgtttc tagtcatcat aatgtaattt gccagttact ttctgactac 1080 aaagaaaaacagatgctaaa agtctcttct gaaaacagca atccaggaaa tgtctcaaga 1140 accagaaataaataagggtg gtgata 1166 <210> SEQ ID NO 340 <211> LENGTH: 384 <212> TYPE:PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 340 Met Val Ala Glu AlaGly Ser Met Pro Ala Ala Ser Ser Val Lys Lys 5 10 15 Pro Phe Gly Leu ArgSer Lys Met Gly Lys Trp Cys Arg His Cys Phe 20 25 30 Pro Trp Cys Arg GlySer Gly Lys Ser Asn Val Gly Thr Ser Gly Asp 35 40 45 His Asp Asp Ser AlaMet Lys Thr Leu Arg Ser Lys Met Gly Lys Trp 50 55 60 Cys Arg His Cys PhePro Trp Cys Arg Gly Ser Ser Lys Ser Asn Val 65 70 75 80 Gly Thr Ser GlyAsp His Asp Asp Ser Ala Met Lys Thr Leu Arg Ser 85 90 95 Lys Met Gly LysTrp Cys Cys His Cys Phe Pro Cys Cys Arg Gly Ser 100 105 110 Gly Lys SerLys Val Gly Pro Trp Gly Asp Tyr Asp Asp Ser Ala Phe 115 120 125 Met GluPro Arg Tyr His Val Arg Arg Glu Asp Leu Asp Lys Leu His 130 135 140 ArgAla Ala Trp Trp Gly Lys Val Pro Arg Lys Asp Leu Ile Val Met 145 150 155160 Leu Lys Asp Thr Asp Met Asn Lys Lys Asp Lys Gln Lys Arg Thr Ala 165170 175 Leu His Leu Ala Ser Ala Asn Gly Asn Ser Glu Val Val Lys Leu Leu180 185 190 Leu Asp Arg Arg Cys Gln Leu Asn Ile Leu Asp Asn Lys Lys ArgThr 195 200 205 Ala Leu Thr Lys Ala Val Gln Cys Arg Glu Asp Glu Cys AlaLeu Met 210 215 220 Leu Leu Glu His Gly Thr Asp Pro Asn Ile Pro Asp GluTyr Gly Asn 225 230 235 240 Thr Ala Leu His Tyr Ala Ile Tyr Asn Glu AspLys Leu Met Ala Lys 245 250 255 Ala Leu Leu Leu Tyr Gly Ala Asp Ile GluSer Lys Asn Lys His Gly 260 265 270 Leu Thr Pro Leu Leu Leu Gly Val HisGlu Gln Lys Gln Gln Val Val 275 280 285 Lys Phe Leu Ile Lys Lys Lys AlaAsn Leu Asn Ala Leu Asp Arg Tyr 290 295 300 Gly Arg Thr Ala Leu Ile LeuAla Val Cys Cys Gly Ser Ala Ser Ile 305 310 315 320 Val Ser Leu Leu LeuGlu Gln Asn Ile Asp Val Ser Ser Gln Asp Leu 325 330 335 Ser Gly Gln ThrAla Arg Glu Tyr Ala Val Ser Ser His His Asn Val 340 345 350 Ile Cys GlnLeu Leu Ser Asp Tyr Lys Glu Lys Gln Met Leu Lys Val 355 360 365 Ser SerGlu Asn Ser Asn Pro Gly Asn Val Ser Arg Thr Arg Asn Lys 370 375 380<210> SEQ ID NO 341 <211> LENGTH: 876 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 341 atgcatcttt catttcctgc atttcttcctccctggatgg acagggggag cggcaagagc 60 aacgtgggca cttctggaga ccacaacgactcctctgtga agacgcttgg gagcaagagg 120 tgcaagtggt gctgccactg cttcccctgctgcaggggga gcggcaagag caacgtggtc 180 gcttggggag actacgatga cagcgccttcatggatccca ggtaccacgt ccatggagaa 240 gatctggaca agctccacag agctgcctggtggggtaaag tccccagaaa ggatctcatc 300 gtcatgctca gggacacgga tgtgaacaagagggacaagc aaaagaggac tgctctacat 360 ctggcctctg ccaatgggaa ttcagaagtagtaaaactcg tgctggacag acgatgtcaa 420 cttaatgtcc ttgacaacaa aaagaggacagctctgacaa aggccgtaca atgccaggaa 480 gatgaatgtg cgttaatgtt gctggaacatggcactgatc caaatattcc agatgagtat 540 ggaaatacca ctctacacta tgctgtctacaatgaagata aattaatggc caaagcactg 600 ctcttatacg gtgctgatat cgaatcaaaaaacaagcatg gcctcacacc actgctactt 660 ggtatacatg agcaaaaaca gcaagtggtgaaatttttaa tcaagaaaaa agcgaattta 720 aatgcgctgg atagatatgg aagaactgctctcatacttg ctgtatgttg tggatcagca 780 agtatagtca gccctctact tgagcaaaatgttgatgtat cttctcaaga tctggaaaga 840 cggccagaga gtatgctgtt tctagtcatcatcatg 876 <210> SEQ ID NO 342 <211> LENGTH: 876 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 342 atgcatcttt catttcctgcatttcttcct ccctggatgg acagggggag cggcaagagc 60 aacgtgggca cttctggagaccacaacgac tcctctgtga agacgcttgg gagcaagagg 120 tgcaagtggt gctgccactgcttcccctgc tgcaggggga gcggcaagag caacgtgggc 180 gcttggggag actacgatgacagcgccttc atggatccca ggtaccacgt ccatggagaa 240 gatctggaca agctccacagagctgcctgg tggggtaaag tccccagaaa ggatctcatc 300 gtcatgctca gggacactgatgtgaacaag agggacaagc aaaagaggac tgctctacat 360 ctggcctctg ccaatgggaattcagaagta gtaaaactcg tgctggacag acgatgtcaa 420 cttaatgtcc ttgacaacaaaaagaggaca gctctgacaa aggccgtaca atgccaggaa 480 gatgaatgtg cgttaatgttgctggaacat ggcactgatc caaatattcc agatgagtat 540 ggaaatacca ctctacactatgctgtctac aatgaagata aattaatggc caaagcactg 600 ctcttatacg gtgctgatatcgaatcaaaa aacaagcatg gcctcacacc actgctactt 660 ggtatacatg agcaaaaacagcaagtggtg aaatttttaa tcaagaaaaa agcgaattta 720 aatgcgctgg atagatatggaagaactgct ctcatacttg ctgtatgttg tggatcagca 780 agtatagtca gccctctacttgagcaaaat gttgatgtat cttctcaaga tctggaaaga 840 cggccagaga gtatgctgtttctagtcatc atcatg 876 <210> SEQ ID NO 343 <211> LENGTH: 933 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 343 atggtggttgaggttgattc aatgccggct gcctcttctg tgaagaagcc atttggtctc 60 aggagcaagatgggcaagtg gtgctgcttt ccctgctgca gggggagcgg caagagcaac 120 gtgggcacttctggagacca caacgactcc tctgtgaaga cgcttgggag caagaggtgc 180 aagtggtgctgccactgctt cccctgctgc agggggagcg gcaagagcaa cgtgggcgct 240 tggggagactacgatgacag cgccttcatg gatcccaggt accacgtcca tggagaagat 300 ctggacaagctccacagagc tgcctggtgg ggtaaagtcc ccagaaagga tctcatcgtc 360 atgctcagggacactgatgt gaacaagagg gacaagcaaa agaggactgc tctacatctg 420 gcctctgccaatgggaattc agaagtagta aaactcgtgc tggacagacg atgtcaactt 480 aatgtccttgacaacaaaaa gaggacagct ctgacaaagg ccgtacaatg ccaggaagat 540 gaatgtgcgttaatgttgct ggaacatggc actgatccaa atattccaga tgagtatgga 600 aataccactctacactatgc tgtctacaat gaagataaat taatggccaa agcactgctc 660 ttatacggtgctgatatcga atcaaaaaac aagcatggcc tcacaccact gctacttggt 720 atacatgagcaaaaacagca agtggtgaaa tttttaatca agaaaaaagc gaatttaaat 780 gcgctggatagatatggaag aactgctctc atacttgctg tatgttgtgg atcagcaagt 840 atagtcagccctctacttga gcaaaatgtt gatgtatctt ctcaagatct ggaaagacgg 900 ccagagagtatgctgtttct agtcatcatc atg 933 <210> SEQ ID NO 344 <211> LENGTH: 939<212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 344atggtggttg aggttgattc aatgccggct gcctcttctg tgaagaagcc atttggtctc 60aggagcaaga tgggcaagtg gtgctgccac tgctttccct gctgcagggg gagcggcaag 120agcaacgtgg gcacttctgg agaccacaac gactcctctg tgaagacgct tgggagcaag 180aggtgcaagt ggtgctgcca ctgcttcccc tgctgcaggg ggagcggcaa gagcaacgtg 240gtcgcttggg gagactacga tgacagcgcc ttcatggatc ccaggtacca cgtccatgga 300gaagatctgg acaagctcca cagagctgcc tggtggggta aagtccccag aaaggatctc 360atcgtcatgc tcagggacac ggatgtgaac aagagggaca agcaaaagag gactgctcta 420catctggcct ctgccaatgg gaattcagaa gtagtaaaac tcgtgctgga cagacgatgt 480caacttaatg tccttgacaa caaaaagagg acagctctga caaaggccgt acaatgccag 540gaagatgaat gtgcgttaat gttgctggaa catggcactg atccaaatat tccagatgag 600tatggaaata ccactctaca ctatgctgtc tacaatgaag ataaattaat ggccaaagca 660ctgctcttat acggtgctga tatcgaatca aaaaacaagc atggcctcac accactgcta 720cttggtatac atgagcaaaa acagcaagtg gtgaaatttt taatcaagaa aaaagcgaat 780ttaaatgcgc tggatagata tggaagaact gctctcatac ttgctgtatg ttgtggatca 840gcaagtatag tcagccctct acttgagcaa aatgttgatg tatcttctca agatctggaa 900agacggccag agagtatgct gtttctagtc atcatcatg 939 <210> SEQ ID NO 345 <211>LENGTH: 292 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:345 Met His Leu Ser Phe Pro Ala Phe Leu Pro Pro Trp Met Asp Arg Gly 5 1015 Ser Gly Lys Ser Asn Val Gly Thr Ser Gly Asp His Asn Asp Ser Ser 20 2530 Val Lys Thr Leu Gly Ser Lys Arg Cys Lys Trp Cys Cys His Cys Phe 35 4045 Pro Cys Cys Arg Gly Ser Gly Lys Ser Asn Val Val Ala Trp Gly Asp 50 5560 Tyr Asp Asp Ser Ala Phe Met Asp Pro Arg Tyr His Val His Gly Glu 65 7075 80 Asp Leu Asp Lys Leu His Arg Ala Ala Trp Trp Gly Lys Val Pro Arg 8590 95 Lys Asp Leu Ile Val Met Leu Arg Asp Thr Asp Val Asn Lys Arg Asp100 105 110 Lys Gln Lys Arg Thr Ala Leu His Leu Ala Ser Ala Asn Gly AsnSer 115 120 125 Glu Val Val Lys Leu Val Leu Asp Arg Arg Cys Gln Leu AsnVal Leu 130 135 140 Asp Asn Lys Lys Arg Thr Ala Leu Thr Lys Ala Val GlnCys Gln Glu 145 150 155 160 Asp Glu Cys Ala Leu Met Leu Leu Glu His GlyThr Asp Pro Asn Ile 165 170 175 Pro Asp Glu Tyr Gly Asn Thr Thr Leu HisTyr Ala Val Tyr Asn Glu 180 185 190 Asp Lys Leu Met Ala Lys Ala Leu LeuLeu Tyr Gly Ala Asp Ile Glu 195 200 205 Ser Lys Asn Lys His Gly Leu ThrPro Leu Leu Leu Gly Ile His Glu 210 215 220 Gln Lys Gln Gln Val Val LysPhe Leu Ile Lys Lys Lys Ala Asn Leu 225 230 235 240 Asn Ala Leu Asp ArgTyr Gly Arg Thr Ala Leu Ile Leu Ala Val Cys 245 250 255 Cys Gly Ser AlaSer Ile Val Ser Pro Leu Leu Glu Gln Asn Val Asp 260 265 270 Val Ser SerGln Asp Leu Glu Arg Arg Pro Glu Ser Met Leu Phe Leu 275 280 285 Val IleIle Met 290 <210> SEQ ID NO 346 <211> LENGTH: 292 <212> TYPE: PRT <213>ORGANISM: Homo sapiens <400> SEQUENCE: 346 Met His Leu Ser Phe Pro AlaPhe Leu Pro Pro Trp Met Asp Arg Gly 5 10 15 Ser Gly Lys Ser Asn Val GlyThr Ser Gly Asp His Asn Asp Ser Ser 20 25 30 Val Lys Thr Leu Gly Ser LysArg Cys Lys Trp Cys Cys His Cys Phe 35 40 45 Pro Cys Cys Arg Gly Ser GlyLys Ser Asn Val Gly Ala Trp Gly Asp 50 55 60 Tyr Asp Asp Ser Ala Phe MetAsp Pro Arg Tyr His Val His Gly Glu 65 70 75 80 Asp Leu Asp Lys Leu HisArg Ala Ala Trp Trp Gly Lys Val Pro Arg 85 90 95 Lys Asp Leu Ile Val MetLeu Arg Asp Thr Asp Val Asn Lys Arg Asp 100 105 110 Lys Gln Lys Arg ThrAla Leu His Leu Ala Ser Ala Asn Gly Asn Ser 115 120 125 Glu Val Val LysLeu Val Leu Asp Arg Arg Cys Gln Leu Asn Val Leu 130 135 140 Asp Asn LysLys Arg Thr Ala Leu Thr Lys Ala Val Gln Cys Gln Glu 145 150 155 160 AspGlu Cys Ala Leu Met Leu Leu Glu His Gly Thr Asp Pro Asn Ile 165 170 175Pro Asp Glu Tyr Gly Asn Thr Thr Leu His Tyr Ala Val Tyr Asn Glu 180 185190 Asp Lys Leu Met Ala Lys Ala Leu Leu Leu Tyr Gly Ala Asp Ile Glu 195200 205 Ser Lys Asn Lys His Gly Leu Thr Pro Leu Leu Leu Gly Ile His Glu210 215 220 Gln Lys Gln Gln Val Val Lys Phe Leu Ile Lys Lys Lys Ala AsnLeu 225 230 235 240 Asn Ala Leu Asp Arg Tyr Gly Arg Thr Ala Leu Ile LeuAla Val Cys 245 250 255 Cys Gly Ser Ala Ser Ile Val Ser Pro Leu Leu GluGln Asn Val Asp 260 265 270 Val Ser Ser Gln Asp Leu Glu Arg Arg Pro GluSer Met Leu Phe Leu 275 280 285 Val Ile Ile Met 290 <210> SEQ ID NO 347<211> LENGTH: 311 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 347 Met Val Val Glu Val Asp Ser Met Pro Ala Ala Ser Ser ValLys Lys 5 10 15 Pro Phe Gly Leu Arg Ser Lys Met Gly Lys Trp Cys Cys PhePro Cys 20 25 30 Cys Arg Gly Ser Gly Lys Ser Asn Val Gly Thr Ser Gly AspHis Asn 35 40 45 Asp Ser Ser Val Lys Thr Leu Gly Ser Lys Arg Cys Lys TrpCys Cys 50 55 60 His Cys Phe Pro Cys Cys Arg Gly Ser Gly Lys Ser Asn ValGly Ala 65 70 75 80 Trp Gly Asp Tyr Asp Asp Ser Ala Phe Met Asp Pro ArgTyr His Val 85 90 95 His Gly Glu Asp Leu Asp Lys Leu His Arg Ala Ala TrpTrp Gly Lys 100 105 110 Val Pro Arg Lys Asp Leu Ile Val Met Leu Arg AspThr Asp Val Asn 115 120 125 Lys Arg Asp Lys Gln Lys Arg Thr Ala Leu HisLeu Ala Ser Ala Asn 130 135 140 Gly Asn Ser Glu Val Val Lys Leu Val LeuAsp Arg Arg Cys Gln Leu 145 150 155 160 Asn Val Leu Asp Asn Lys Lys ArgThr Ala Leu Thr Lys Ala Val Gln 165 170 175 Cys Gln Glu Asp Glu Cys AlaLeu Met Leu Leu Glu His Gly Thr Asp 180 185 190 Pro Asn Ile Pro Asp GluTyr Gly Asn Thr Thr Leu His Tyr Ala Val 195 200 205 Tyr Asn Glu Asp LysLeu Met Ala Lys Ala Leu Leu Leu Tyr Gly Ala 210 215 220 Asp Ile Glu SerLys Asn Lys His Gly Leu Thr Pro Leu Leu Leu Gly 225 230 235 240 Ile HisGlu Gln Lys Gln Gln Val Val Lys Phe Leu Ile Lys Lys Lys 245 250 255 AlaAsn Leu Asn Ala Leu Asp Arg Tyr Gly Arg Thr Ala Leu Ile Leu 260 265 270Ala Val Cys Cys Gly Ser Ala Ser Ile Val Ser Pro Leu Leu Glu Gln 275 280285 Asn Val Asp Val Ser Ser Gln Asp Leu Glu Arg Arg Pro Glu Ser Met 290295 300 Leu Phe Leu Val Ile Ile Met 305 310 <210> SEQ ID NO 348 <211>LENGTH: 313 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:348 Met Val Val Glu Val Asp Ser Met Pro Ala Ala Ser Ser Val Lys Lys 5 1015 Pro Phe Gly Leu Arg Ser Lys Met Gly Lys Trp Cys Cys His Cys Phe 20 2530 Pro Cys Cys Arg Gly Ser Gly Lys Ser Asn Val Gly Thr Ser Gly Asp 35 4045 His Asn Asp Ser Ser Val Lys Thr Leu Gly Ser Lys Arg Cys Lys Trp 50 5560 Cys Cys His Cys Phe Pro Cys Cys Arg Gly Ser Gly Lys Ser Asn Val 65 7075 80 Val Ala Trp Gly Asp Tyr Asp Asp Ser Ala Phe Met Asp Pro Arg Tyr 8590 95 His Val His Gly Glu Asp Leu Asp Lys Leu His Arg Ala Ala Trp Trp100 105 110 Gly Lys Val Pro Arg Lys Asp Leu Ile Val Met Leu Arg Asp ThrAsp 115 120 125 Val Asn Lys Arg Asp Lys Gln Lys Arg Thr Ala Leu His LeuAla Ser 130 135 140 Ala Asn Gly Asn Ser Glu Val Val Lys Leu Val Leu AspArg Arg Cys 145 150 155 160 Gln Leu Asn Val Leu Asp Asn Lys Lys Arg ThrAla Leu Thr Lys Ala 165 170 175 Val Gln Cys Gln Glu Asp Glu Cys Ala LeuMet Leu Leu Glu His Gly 180 185 190 Thr Asp Pro Asn Ile Pro Asp Glu TyrGly Asn Thr Thr Leu His Tyr 195 200 205 Ala Val Tyr Asn Glu Asp Lys LeuMet Ala Lys Ala Leu Leu Leu Tyr 210 215 220 Gly Ala Asp Ile Glu Ser LysAsn Lys His Gly Leu Thr Pro Leu Leu 225 230 235 240 Leu Gly Ile His GluGln Lys Gln Gln Val Val Lys Phe Leu Ile Lys 245 250 255 Lys Lys Ala AsnLeu Asn Ala Leu Asp Arg Tyr Gly Arg Thr Ala Leu 260 265 270 Ile Leu AlaVal Cys Cys Gly Ser Ala Ser Ile Val Ser Pro Leu Leu 275 280 285 Glu GlnAsn Val Asp Val Ser Ser Gln Asp Leu Glu Arg Arg Pro Glu 290 295 300 SerMet Leu Phe Leu Val Ile Ile Met 305 310 <210> SEQ ID NO 349 <211>LENGTH: 30 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:349 Val Asn Lys Lys Asp Lys Gln Lys Arg Thr Ala Leu His Leu Ala Ser 1 510 15 Ala Asn Gly Asn Ser Glu Val Val Lys Leu Leu Leu Asp Arg 20 25 30<210> SEQ ID NO 350 <211> LENGTH: 30 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 350 Ala Leu His Leu Ala Ser Ala Asn Gly AsnSer Glu Val Val Lys Leu 1 5 10 15 Leu Leu Asp Arg Arg Cys Gln Leu AsnVal Leu Asp Asn Lys 20 25 30 <210> SEQ ID NO 351 <211> LENGTH: 25 <212>TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 351 Gly Ser AlaSer Ile Val Ser Leu Leu Leu Glu Gln Asn Ile Asp Val 1 5 10 15 Ser SerGln Asp Leu Ser Gly Gln Thr 20 25 <210> SEQ ID NO 352 <211> LENGTH: 20<212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 352 Val ValGlu Val Asp Ser Met Pro Ala Ala Ser Ser Val Lys Lys Pro 1 5 10 15 PheGly Leu Arg 20 <210> SEQ ID NO 353 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 353 Ser Met Pro Ala Ala SerSer Val Lys Lys Pro Phe Gly Leu Arg Ser 1 5 10 15 Lys Met Gly Lys 20<210> SEQ ID NO 354 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 354 Ser Ser Val Lys Lys Pro Phe Gly Leu ArgSer Lys Met Gly Lys Trp 1 5 10 15 Cys Cys Arg Cys 20 <210> SEQ ID NO 355<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 355 Pro Phe Gly Leu Arg Ser Lys Met Gly Lys Trp Cys Cys ArgCys Phe 1 5 10 15 Pro Cys Cys Arg 20 <210> SEQ ID NO 356 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 356 SerLys Met Gly Lys Trp Cys Cys Arg Cys Phe Pro Cys Cys Arg Glu 1 5 10 15Ser Gly Lys Ser 20 <210> SEQ ID NO 357 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 357 Trp Cys Cys Arg Cys PhePro Cys Cys Arg Glu Ser Gly Lys Ser Asn 1 5 10 15 Val Gly Thr Ser 20<210> SEQ ID NO 358 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 358 Phe Pro Cys Cys Arg Glu Ser Gly Lys SerAsn Val Gly Thr Ser Gly 1 5 10 15 Asp His Asp Asp 20 <210> SEQ ID NO 359<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 359 Glu Ser Gly Lys Ser Asn Val Gly Thr Ser Gly Asp His AspAsp Ser 1 5 10 15 Ala Met Lys Thr 20 <210> SEQ ID NO 360 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 360 AsnVal Gly Thr Ser Gly Asp His Asp Asp Ser Ala Met Lys Thr Leu 1 5 10 15Arg Ser Lys Met 20 <210> SEQ ID NO 361 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 361 Gly Asp His Asp Asp SerAla Met Lys Thr Leu Arg Ser Lys Met Gly 1 5 10 15 Lys Trp Cys Arg 20<210> SEQ ID NO 362 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 362 Ser Ala Met Lys Thr Leu Arg Ser Lys MetGly Lys Trp Cys Arg His 1 5 10 15 Cys Phe Pro Cys 20 <210> SEQ ID NO 363<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 363 Leu Arg Ser Lys Met Gly Lys Trp Cys Arg His Cys Phe ProCys Cys 1 5 10 15 Arg Gly Ser Gly 20 <210> SEQ ID NO 364 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 364 GlyLys Trp Cys Arg His Cys Phe Pro Cys Cys Arg Gly Ser Gly Lys 1 5 10 15Ser Asn Val Gly 20 <210> SEQ ID NO 365 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 365 His Cys Phe Pro Cys CysArg Gly Ser Gly Lys Ser Asn Val Gly Ala 1 5 10 15 Ser Gly Asp His 20<210> SEQ ID NO 366 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 366 Cys Arg Gly Ser Gly Lys Ser Asn Val GlyAla Ser Gly Asp His Asp 1 5 10 15 Asp Ser Ala Met 20 <210> SEQ ID NO 367<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 367 Lys Ser Asn Val Gly Ala Ser Gly Asp His Asp Asp Ser AlaMet Lys 1 5 10 15 Thr Leu Arg Asn 20 <210> SEQ ID NO 368 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 368 AlaSer Gly Asp His Asp Asp Ser Ala Met Lys Thr Leu Arg Asn Lys 1 5 10 15Met Gly Lys Trp 20 <210> SEQ ID NO 369 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 369 Asp Asp Ser Ala Met LysThr Leu Arg Asn Lys Met Gly Lys Trp Cys 1 5 10 15 Cys His Cys Phe 20<210> SEQ ID NO 370 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 370 Lys Thr Leu Arg Asn Lys Met Gly Lys TrpCys Cys His Cys Phe Pro 1 5 10 15 Cys Cys Arg Gly 20 <210> SEQ ID NO 371<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 371 Lys Met Gly Lys Trp Cys Cys His Cys Phe Pro Cys Cys ArgGly Ser 1 5 10 15 Gly Lys Ser Lys 20 <210> SEQ ID NO 372 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 372 CysCys His Cys Phe Pro Cys Cys Arg Gly Ser Gly Lys Ser Lys Val 1 5 10 15Gly Ala Trp Gly 20 <210> SEQ ID NO 373 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 373 Pro Cys Cys Arg Gly SerGly Lys Ser Lys Val Gly Ala Trp Gly Asp 1 5 10 15 Tyr Asp Asp Ser 20<210> SEQ ID NO 374 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 374 Ser Gly Lys Ser Lys Val Gly Ala Trp GlyAsp Tyr Asp Asp Ser Ala 1 5 10 15 Phe Met Glu Pro 20 <210> SEQ ID NO 375<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 375 Val Gly Ala Trp Gly Asp Tyr Asp Asp Ser Ala Phe Met GluPro Arg 1 5 10 15 Tyr His Val Arg 20 <210> SEQ ID NO 376 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 376 AspTyr Asp Asp Ser Ala Phe Met Glu Pro Arg Tyr His Val Arg Gly 1 5 10 15Glu Asp Leu Asp 20 <210> SEQ ID NO 377 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 377 Ala Phe Met Glu Pro ArgTyr His Val Arg Gly Glu Asp Leu Asp Lys 1 5 10 15 Leu His Arg Ala 20<210> SEQ ID NO 378 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 378 Arg Tyr His Val Arg Gly Glu Asp Leu AspLys Leu His Arg Ala Ala 1 5 10 15 Trp Trp Gly Lys 20 <210> SEQ ID NO 379<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 379 Gly Glu Asp Leu Asp Lys Leu His Arg Ala Ala Trp Trp GlyLys Val 1 5 10 15 Pro Arg Lys Asp 20 <210> SEQ ID NO 380 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 380 LysLeu His Arg Ala Ala Trp Trp Gly Lys Val Pro Arg Lys Asp Leu 1 5 10 15Ile Val Met Leu 20 <210> SEQ ID NO 381 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 381 Ala Trp Trp Gly Lys ValPro Arg Lys Asp Leu Ile Val Met Leu Arg 1 5 10 15 Asp Thr Asp Val 20<210> SEQ ID NO 382 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 382 Val Pro Arg Lys Asp Leu Ile Val Met LeuArg Asp Thr Asp Val Asn 1 5 10 15 Lys Lys Asp Lys 20 <210> SEQ ID NO 383<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 383 Leu Ile Val Met Leu Arg Asp Thr Asp Val Asn Lys Lys AspLys Gln 1 5 10 15 Lys Arg Thr Ala 20 <210> SEQ ID NO 384 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 384 ArgAsp Thr Asp Val Asn Lys Lys Asp Lys Gln Lys Arg Thr Ala Leu 1 5 10 15His Leu Ala Ser 20 <210> SEQ ID NO 385 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 385 Asn Lys Lys Asp Lys GlnLys Arg Thr Ala Leu His Leu Ala Ser Ala 1 5 10 15 Asn Gly Asn Ser 20<210> SEQ ID NO 386 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 386 Gln Lys Arg Thr Ala Leu His Leu Ala SerAla Asn Gly Asn Ser Glu 1 5 10 15 Val Val Lys Leu 20 <210> SEQ ID NO 387<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 387 Leu His Leu Ala Ser Ala Asn Gly Asn Ser Glu Val Val LysLeu Leu 1 5 10 15 Leu Asp Arg Arg 20 <210> SEQ ID NO 388 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 388 AlaAsn Gly Asn Ser Glu Val Val Lys Leu Leu Leu Asp Arg Arg Cys 1 5 10 15Gln Leu Asn Val 20 <210> SEQ ID NO 389 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 389 Glu Val Val Lys Leu LeuLeu Asp Arg Arg Cys Gln Leu Asn Val Leu 1 5 10 15 Asp Asn Lys Lys 20<210> SEQ ID NO 390 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 390 Leu Leu Asp Arg Arg Cys Gln Leu Asn ValLeu Asp Asn Lys Lys Arg 1 5 10 15 Thr Ala Leu Ile 20 <210> SEQ ID NO 391<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 391 Cys Gln Leu Asn Val Leu Asp Asn Lys Lys Arg Thr Ala LeuIle Lys 1 5 10 15 Ala Val Gln Cys 20 <210> SEQ ID NO 392 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 392 LeuAsp Asn Lys Lys Arg Thr Ala Leu Ile Lys Ala Val Gln Cys Gln 1 5 10 15Glu Asp Glu Cys 20 <210> SEQ ID NO 393 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 393 Arg Thr Ala Leu Ile LysAla Val Gln Cys Gln Glu Asp Glu Cys Ala 1 5 10 15 Leu Met Leu Leu 20<210> SEQ ID NO 394 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 394 Lys Ala Val Gln Cys Gln Glu Asp Glu CysAla Leu Met Leu Leu Glu 1 5 10 15 His Gly Thr Asp 20 <210> SEQ ID NO 395<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 395 Gln Glu Asp Glu Cys Ala Leu Met Leu Leu Glu His Gly ThrAsp Pro 1 5 10 15 Asn Ile Pro Asp 20 <210> SEQ ID NO 396 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 396 AlaLeu Met Leu Leu Glu His Gly Thr Asp Pro Asn Ile Pro Asp Glu 1 5 10 15Tyr Gly Asn Thr 20 <210> SEQ ID NO 397 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 397 Glu His Gly Thr Asp ProAsn Ile Pro Asp Glu Tyr Gly Asn Thr Thr 1 5 10 15 Leu His Tyr Ala 20<210> SEQ ID NO 398 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 398 Pro Asn Ile Pro Asp Glu Tyr Gly Asn ThrThr Leu His Tyr Ala Ile 1 5 10 15 Tyr Asn Glu Asp 20 <210> SEQ ID NO 399<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 399 Glu Tyr Gly Asn Thr Thr Leu His Tyr Ala Ile Tyr Asn GluAsp Lys 1 5 10 15 Leu Met Ala Lys 20 <210> SEQ ID NO 400 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 400 ThrLeu His Tyr Ala Ile Tyr Asn Glu Asp Lys Leu Met Ala Lys Ala 1 5 10 15Leu Leu Leu Tyr 20 <210> SEQ ID NO 401 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 401 Ile Tyr Asn Glu Asp LysLeu Met Ala Lys Ala Leu Leu Leu Tyr Gly 1 5 10 15 Ala Asp Ile Glu 20<210> SEQ ID NO 402 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 402 Lys Leu Met Ala Lys Ala Leu Leu Leu TyrGly Ala Asp Ile Glu Ser 1 5 10 15 Lys Asn Lys His 20 <210> SEQ ID NO 403<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 403 Ala Leu Leu Leu Tyr Gly Ala Asp Ile Glu Ser Lys Asn LysHis Gly 1 5 10 15 Leu Thr Pro Leu 20 <210> SEQ ID NO 404 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 404 GlyAla Asp Ile Glu Ser Lys Asn Lys His Gly Leu Thr Pro Leu Leu 1 5 10 15Leu Gly Val His 20 <210> SEQ ID NO 405 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 405 Ser Lys Asn Lys His GlyLeu Thr Pro Leu Leu Leu Gly Val His Glu 1 5 10 15 Gln Lys Gln Gln 20<210> SEQ ID NO 406 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 406 Gly Leu Thr Pro Leu Leu Leu Gly Val HisGlu Gln Lys Gln Gln Val 1 5 10 15 Val Lys Phe Leu 20 <210> SEQ ID NO 407<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 407 Leu Leu Gly Val His Glu Gln Lys Gln Gln Val Val Lys PheLeu Ile 1 5 10 15 Lys Lys Lys Ala 20 <210> SEQ ID NO 408 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 408 GluGln Lys Gln Gln Val Val Lys Phe Leu Ile Lys Lys Lys Ala Asn 1 5 10 15Leu Asn Ala Leu 20 <210> SEQ ID NO 409 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 409 Val Val Lys Phe Leu IleLys Lys Lys Ala Asn Leu Asn Ala Leu Asp 1 5 10 15 Arg Tyr Gly Arg 20<210> SEQ ID NO 410 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 410 Ile Lys Lys Lys Ala Asn Leu Asn Ala LeuAsp Arg Tyr Gly Thr Arg 1 5 10 15 Ala Leu Ile Leu 20 <210> SEQ ID NO 411<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 411 Asn Leu Asn Ala Leu Asp Arg Tyr Gly Arg Thr Ala Leu IleLeu Ala 1 5 10 15 Val Cys Cys Gly 20 <210> SEQ ID NO 412 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 412 AspArg Tyr Gly Arg Thr Ala Leu Ile Leu Ala Val Cys Cys Gly Ser 1 5 10 15Ala Ser Ile Val 20 <210> SEQ ID NO 413 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 413 Thr Ala Leu Ile Leu AlaVal Cys Cys Gly Ser Ala Ser Ile Val Ser 1 5 10 15 Leu Leu Leu Glu 20<210> SEQ ID NO 414 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 414 Ala Val Cys Cys Gly Ser Ala Ser Ile ValSer Leu Leu Leu Glu Gln 1 5 10 15 Asn Ile Asp Val 20 <210> SEQ ID NO 415<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 415 Ser Ala Ser Ile Val Ser Leu Leu Leu Glu Gln Asn Ile AspVal Ser 1 5 10 15 Ser Gln Asp Leu 20 <210> SEQ ID NO 416 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 416 SerLeu Leu Leu Glu Gln Asn Ile Asp Val Ser Ser Gln Asp Leu Ser 1 5 10 15Gly Gln Thr Ala 20 <210> SEQ ID NO 417 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 417 Gln Asn Ile Asp Val SerSer Gln Asp Leu Ser Gly Gln Thr Ala Arg 1 5 10 15 Glu Tyr Ala Val 20<210> SEQ ID NO 418 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 418 Ser Ser Gln Asp Leu Ser Gly Gln Thr AlaArg Glu Tyr Ala Val Ser 1 5 10 15 Ser His His His 20 <210> SEQ ID NO 419<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 419 Ser Gly Gln Thr Ala Arg Glu Tyr Ala Val Ser Ser His HisHis Val 1 5 10 15 Ile Cys Gln Leu 20 <210> SEQ ID NO 420 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 420 ArgGlu Tyr Ala Val Ser Ser His His His Val Ile Cys Gln Leu Leu 1 5 10 15Ser Asp Tyr Lys 20 <210> SEQ ID NO 421 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 421 Ser Ser His His His ValIle Cys Gln Leu Leu Ser Asp Tyr Lys Glu 1 5 10 15 Lys Gln Met Leu 20<210> SEQ ID NO 422 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 422 Val Ile Cys Gln Leu Leu Ser Asp Tyr LysGlu Lys Gln Met Leu Lys 1 5 10 15 Ile Ser Ser Glu 20 <210> SEQ ID NO 423<211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400>SEQUENCE: 423 Leu Ser Asp Tyr Lys Glu Lys Gln Met Leu Lys Ile Ser SerGlu Asn 1 5 10 15 Ser Asn Pro Glu 20 <210> SEQ ID NO 424 <211> LENGTH:20 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 424 GluLys Gln Met Leu Lys Ile Ser Ser Glu Asn Ser Asn Pro Glu Asn 1 5 10 15Val Ser Arg Thr 20 <210> SEQ ID NO 425 <211> LENGTH: 20 <212> TYPE: PRT<213> ORGANISM: Homo sapiens <400> SEQUENCE: 425 Met Leu Lys Ile Ser SerGlu Asn Ser Asn Pro Glu Asn Val Ser Arg 1 5 10 15 Thr Arg Asn Lys 20<210> SEQ ID NO 426 <211> LENGTH: 33 <212> TYPE: PRT <213> ORGANISM:Homo sapiens <400> SEQUENCE: 426 Ser Lys Met Gly Lys Trp Cys Cys Arg CysPhe Pro Cys Cys Arg Glu 1 5 10 15 Ser Gly Lys Ser Asn Val Gly Thr SerGly Asp His Asp Asp Ser Ala 20 25 30 Met <210> SEQ ID NO 427 <211>LENGTH: 33 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:427 Ser Lys Met Gly Lys Trp Cys Arg His Cys Phe Pro Cys Cys Arg Gly 1 510 15 Ser Gly Lys Ser Asn Val Gly Ala Ser Gly Asp His Asp Asp Ser Ala 2025 30 Met <210> SEQ ID NO 428 <211> LENGTH: 33 <212> TYPE: PRT <213>ORGANISM: Homo sapiens <400> SEQUENCE: 428 Asn Lys Met Gly Lys Trp CysCys His Cys Phe Pro Cys Cys Arg Gly 1 5 10 15 Ser Gly Lys Ser Lys ValGly Ala Trp Gly Asp Tyr Asp Asp Ser Ala 20 25 30 Phe

What is claimed:
 1. An isolated polynucleotide comprising a sequenceselected from the group consisting of: (a) sequences provided in SEQ IDNO: 341-344; (b) complements of the sequences provided in SEQ ID NO:341-344; (c) sequences consisting of at least 20 contiguous residues ofa sequence provided in SEQ ID NO: 341-344; (d) sequences that hybridizeto a sequence provided in SEQ ID NO: 341-344, under highly stringentconditions; (e) sequences having at least 75% identity to a sequence ofSEQ ID NO: 341-344; (f) sequences having at least 90% identity to asequence of SEQ ID NO: 341-344; and (g) degenerate variants of asequence provided in SEQ ID NO: 341-344.
 2. An isolated polypeptidecomprising an amino acid sequence selected from the group consisting of:(a) sequences encoded by a polynucleotide of claim 1; and (b) sequenceshaving at least 70% identity to a sequence encoded by a polynucleotideof claim 1; and (c) sequences having at least 90% identity to a sequenceencoded by a polynucleotide of claim 1; (d) sequences set forth in SEQID NO: 345-428; (e) sequences having at least 70% identity to a sequenceset forth in SEQ ID NO: 345-428; and (f) sequences having at least 90%identity to a sequence set forth in SEQ ID NO: 345-428.
 3. An expressionvector comprising a polynucleotide of claim 1 operably linked to anexpression control sequence.
 4. A host cell transformed or transfectedwith an expression vector according to claim
 3. 5. An isolated antibody,or antigen-binding fragment thereof, that specifically binds to apolypeptide of claim
 2. 6. A method for detecting the presence of acancer in a patient, comprising the steps of: (a) obtaining a biologicalsample from the patient; (b) contacting the biological sample with abinding agent that binds to a polypeptide of claim 2; (c) detecting inthe sample an amount of polypeptide that binds to the binding agent; and(d) comparing the amount of polypeptide to a predetermined cut-off valueand therefrom determining the presence of a cancer in the patient.
 7. Afusion protein comprising at least one polypeptide according to claim 2.8. An oligonucleotide that hybridizes to a sequence recited in SEQ IDNO: 341-344 under highly stringent conditions.
 9. A method forstimulating and/or expanding T cells specific for a tumor protein,comprising contacting T cells with at least one component selected fromthe group consisting of: (a) polypeptides according to claim 2; (b)polynucleotides according to claim 1; and (c) antigen-presenting cellsthat express a polynucleotide according to claim 1, under conditions andfor a time sufficient to permit the stimulation and/or expansion of Tcells.
 10. An isolated T cell population, comprising T cells preparedaccording to the method of claim
 9. 11. A composition comprising a firstcomponent selected from the group consisting of physiologicallyacceptable carriers and immunostimulants, and a second componentselected from the group consisting of: (a) polypeptides according toclaim 2; (b) polynucleotides according to claim 1; (c) antibodiesaccording to claim 5; (d) fusion proteins according to claim 7; (e) Tcell populations according to claim 10; and (f) antigen presenting cellsthat express a polypeptide according to claim
 2. 12. A method forstimulating an immune response in a patient, comprising administering tothe patient a composition of claim
 11. 13. A method for the treatment ofa breast cancer in a patient, comprising administering to the patient acomposition of claim
 11. 14. A method for determining the presence of acancer in a patient, comprising the steps of: (a) obtaining a biologicalsample from the patient; (b) contacting the biological sample with anoligonucleotide according to claim 8; (c) detecting in the sample anamount of a polynucleotide that hybridizes to the oligonucleotide; and(d) comparing the amount of polynucleotide that hybridizes to theoligonucleotide to a predetermined cut-off value, and therefromdetermining the presence of the cancer in the patient.
 15. A diagnostickit comprising at least one oligonucleotide according to claim
 8. 16. Adiagnostic kit comprising at least one antibody according to claim 5 anda detection reagent, wherein the detection reagent comprises a reportergroup.
 17. A method for the treatment of breast cancer in a patient,comprising the steps of: (a) incubating CD4+ and/or CD8+ T cellsisolated from a patient with at least one component selected from thegroup consisting of: (i) polypeptides according to claim 2; (ii)polynucleotides according to claim 1; and (iii) antigen presenting cellsthat express a polypeptide of claim 2, such that T cell proliferate; (b)administering to the patient an effective amount of the proliferated Tcells, and thereby inhibiting the development of a cancer in thepatient.